{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T04:13:15Z","timestamp":1792037595276,"version":"4.4.0"},"reference-count":99,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,5,25]],"date-time":"2022-05-25T00:00:00Z","timestamp":1653436800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Mol. Biosci."],"abstract":"<jats:p>Even though there are a great number of possible conformational states, how a protein generated as a linear unfolded polypeptide efficiently folds into its physiologically active form remained a fascinating and unanswered enigma inside crowded conditions of cells. In this study, various spectroscopic techniques have been exploited to know and understand the effect and mechanism of action of two different sizes of polyethylene glycols, or PEGs (molecular mass \u223c10 and \u223c20\u00a0kilo Daltons, kDa), on cytochrome <jats:italic>c<\/jats:italic> (cyt <jats:italic>c<\/jats:italic>). The outcomes showed that small size of the PEG leads to perturbation of the protein structure, and conversely, large size of the PEG has stabilizing effect on cyt <jats:italic>c<\/jats:italic>. Moreover, binding measurements showed that small size of PEG interacts strongly <jats:italic>via<\/jats:italic> soft interactions compared to the larger size of PEG, the latter being governed more by excluded volume effect or preferential exclusion from the protein. Overall, this finding suggests that conformations of protein may be influenced in cellular crowded conditions <jats:italic>via<\/jats:italic> interactions which depend upon the size of molecule in the environment. This study proposes that both volume exclusion and soft (chemical) interactions governs the protein\u2019s conformation and functional activities. The cellular environment\u2019s internal architecture as evident from crowder size and shape in this study has a significant role.<\/jats:p>","DOI":"10.3389\/fmolb.2022.849683","type":"journal-article","created":{"date-parts":[[2022,5,26]],"date-time":"2022-05-26T17:19:41Z","timestamp":1653585581000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":18,"title":["Size-Dependent Interplay of Volume Exclusion Versus Soft Interactions: Cytochrome c in Macromolecular Crowded Environment"],"prefix":"10.3389","volume":"9","author":[{"given":"Zahoor Ahmad","family":"Parray","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Faizan","family":"Ahmad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anis Ahmad","family":"Chaudhary","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hassan Ahmad","family":"Rudayni","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammed","family":"Al-Zharani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md. Imtaiyaz","family":"Hassan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Asimul","family":"Islam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,5,25]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1329","DOI":"10.1073\/pnas.012458999","article-title":"Conformational Landscape of Cytochrome C Folding Studied by Microsecond-Resolved Small-Angle X-ray Scattering","volume":"99","author":"Akiyama","year":"2002","journal-title":"Proc. Natl. Acad. Sci."},{"key":"B2","doi-asserted-by":"publisher","first-page":"151","DOI":"10.1007\/s12551-016-0198-6","article-title":"UV-Vis Spectroscopy of Tyrosine Side-Groups in Studies of Protein Structure. Part 1: Basic Principles and Properties of Tyrosine Chromophore","volume":"8","author":"Antosiewicz","year":"2016","journal-title":"Biophys. Rev."},{"key":"B3","volume-title":"Discovery Studio Modeling Environment","author":"Biovia","year":"2015"},{"key":"B4","doi-asserted-by":"publisher","first-page":"4316","DOI":"10.1021\/acsomega.7b01864","article-title":"Mixed Macromolecular Crowding: A Protein and Solvent Perspective","volume":"3","author":"Biswas","year":"2018","journal-title":"ACS Omega"},{"key":"B5","doi-asserted-by":"publisher","first-page":"087803","DOI":"10.1103\/PhysRevLett.96.087803","article-title":"Light Scattering and Phase Behavior of Lysozyme-Poly(Ethylene Glycol) Mixtures","volume":"96","author":"Bloustine","year":"2006","journal-title":"Phys. Rev. Lett."},{"key":"B6","doi-asserted-by":"publisher","first-page":"1713","DOI":"10.1016\/s0006-3495(94)80645-9","article-title":"Stereodynamic Properties of the Cooperative Homodimeric Scapharca Inaequivalvis Hemoglobin Studied through Optical Absorption Spectroscopy and Ligand Rebinding Kinetics","volume":"67","author":"Boffi","year":"1994","journal-title":"Biophysical J."},{"key":"B7","doi-asserted-by":"publisher","first-page":"1239","DOI":"10.1007\/s10541-005-0070-y","article-title":"Biochemical Effects of Molecular Crowding","volume":"69","author":"Chebotareva","year":"2004","journal-title":"Biochemistry (Moscow)."},{"key":"B8","doi-asserted-by":"publisher","first-page":"4","DOI":"10.1016\/j.abb.2007.05.014","article-title":"Protein Folding: Then and Now","volume":"469","author":"Chen","year":"2008","journal-title":"Arch. Biochem. Biophys."},{"key":"B9","doi-asserted-by":"publisher","first-page":"304","DOI":"10.1038\/254304a0","article-title":"Structural Invariants in Protein Folding","volume":"254","author":"Chothia","year":"1975","journal-title":"Nature."},{"key":"B10","doi-asserted-by":"publisher","first-page":"685","DOI":"10.1002\/cbic.200700603","article-title":"NMR Spectroscopy Reveals Cytochromec\u2013Poly(Ethylene Glycol) Interactions","volume":"9","author":"Crowley","year":"2008","journal-title":"Chembiochem."},{"key":"B11","doi-asserted-by":"publisher","first-page":"243","DOI":"10.1007\/978-1-4939-2269-7_19","article-title":"Small-Molecule Library Screening by Docking with PyRx","volume":"1263","author":"Dallakyan","year":"2015","journal-title":"Methods Mol. Biol."},{"key":"B12","doi-asserted-by":"publisher","first-page":"843","DOI":"10.1016\/j.ijbiomac.2019.09.029","article-title":"Size-Dependent Macromolecular Crowding Effect on the Thermodynamics of Protein Unfolding Revealed at the Single Molecular Level","volume":"141","author":"Das","year":"2019","journal-title":"Int. J. Biol. Macromol."},{"key":"B13","doi-asserted-by":"publisher","first-page":"5858","DOI":"10.1021\/acs.jpcb.0c03897","article-title":"Shape-Dependent Macromolecular Crowding on the Thermodynamics and Microsecond Conformational Dynamics of Protein Unfolding Revealed at the Single-Molecule Level","volume":"124","author":"Das","year":"2020","journal-title":"J. Phys. Chem. B."},{"key":"B14","doi-asserted-by":"publisher","first-page":"5431","DOI":"10.1021\/bi00071a019","article-title":"Redesign of the interior Hydrophilic Region of Mitochondrial Cytochrome C by Site-Directed Mutagenesis","volume":"32","author":"Davies","year":"1993","journal-title":"Biochemistry"},{"key":"B15","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1098\/rspb.1931.0066","article-title":"The Absorption Spectrum of the Component C of Cytochrome","volume":"109","author":"Dixon","year":"1931","journal-title":"Proc. R. Soc. Lond. Ser. B, Containing Pap. a Biol."},{"key":"B16","doi-asserted-by":"publisher","first-page":"144","DOI":"10.3390\/ijms17020144","article-title":"Insights into Protein-Ligand Interactions: Mechanisms, Models, and Methods","volume":"17","author":"Du","year":"2016","journal-title":"Int. J. Mol. Sci."},{"key":"B17","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/978-0-387-39975-1_1","article-title":"Protein Misassembly: Macromolecular Crowding and Molecular Chaperones","volume":"594","author":"Ellis","year":"2007","journal-title":"Adv. Exp. Med. Biol."},{"key":"B18","doi-asserted-by":"publisher","first-page":"114","DOI":"10.1016\/s0959-440x(00)00172-x","article-title":"Macromolecular Crowding: An Important but Neglected Aspect of the Intracellular Environment","volume":"11","author":"Ellis","year":"2001","journal-title":"Curr. Opin. Struct. Biol."},{"key":"B19","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1016\/j.bbapap.2004.09.013","article-title":"The Heme Iron Coordination of Unfolded Ferric and Ferrous Cytochrome C in Neutral and Acidic Urea Solutions. Spectroscopic and Electrochemical Studies","volume":"1703","author":"Fedurco","year":"2004","journal-title":"Biochim. Biophys. Acta (Bba) - Proteins Proteomics"},{"key":"B20","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1016\/s0091-679x(07)84004-0","article-title":"Isothermal Titration Calorimetry: Experimental Design, Data Analysis, and Probing Macromolecule\/Ligand Binding and Kinetic Interactions","volume":"84","author":"Freyer","year":"2008","journal-title":"Methods Cell Biol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"957","DOI":"10.1074\/jbc.m006861200","article-title":"Excluded Volume Effects on the Refolding and Assembly of an Oligomeric Protein","volume":"276","author":"Gal\u00e1n","year":"2001","journal-title":"J. Biol. Chem."},{"key":"B22","doi-asserted-by":"publisher","first-page":"291","DOI":"10.1002\/bmb.20636","article-title":"Illustrating the Machinery of Life: Viruses","volume":"40","author":"Goodsell","year":"2012","journal-title":"Biochem. Mol. Biol. Educ."},{"key":"B23","doi-asserted-by":"publisher","first-page":"3480","DOI":"10.1021\/bi00466a009","article-title":"Mechanism of Acid-Induced Folding of Proteins","volume":"29","author":"Goto","year":"1990","journal-title":"Biochemistry"},{"key":"B24","doi-asserted-by":"publisher","first-page":"4108","DOI":"10.1021\/bi00838a031","article-title":"Computed Circular Dichroism Spectra for the Evaluation of Protein Conformation","volume":"8","author":"Greenfield","year":"1969","journal-title":"Biochemistry"},{"key":"B25","doi-asserted-by":"publisher","first-page":"1658","DOI":"10.1073\/pnas.54.6.1658","article-title":"Complex Formation between Methionine and a Heme Peptide from Cytochrome C","volume":"54","author":"Harbury","year":"1965","journal-title":"Proc. Natl. Acad. Sci."},{"key":"B26","doi-asserted-by":"publisher","first-page":"15160","DOI":"10.1016\/s0021-9258(18)77236-0","article-title":"\u201cMacromolecular Crowding\u201d: Thermodynamic Consequences for Protein-Protein Interactions within the T4 DNA Replication Complex","volume":"265","author":"Jarvis","year":"1990","journal-title":"J. Biol. Chem."},{"key":"B27","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1038\/nchembio727","article-title":"Cytochrome C Acts as a Cardiolipin Oxygenase Required for Release of Proapoptotic Factors","volume":"1","author":"Kagan","year":"2005","journal-title":"Nat. Chem. Biol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"349","DOI":"10.2174\/1389203003381315","article-title":"The Use of Circular Dichroism in the Investigation of Protein Structure and Function","volume":"1","author":"Kelly","year":"2000","journal-title":"Cpps."},{"key":"B29","doi-asserted-by":"publisher","first-page":"327","DOI":"10.1039\/c0mt00078g","article-title":"Conformation and Thermodynamic Stability of Pre-Molten and Molten Globule States of Mammalian Cytochromes-C","volume":"3","author":"Khan","year":"2011","journal-title":"Metallomics."},{"key":"B30","doi-asserted-by":"publisher","first-page":"202","DOI":"10.1016\/j.ijbiomac.2016.06.096","article-title":"Structural and Thermodynamic Characterisation of L94F Mutant of Horse Cytochrome C","volume":"92","author":"Khan","year":"2016","journal-title":"Int. J. Biol. Macromolecules"},{"key":"B31","doi-asserted-by":"publisher","first-page":"2139","DOI":"10.1529\/biophysj.106.097717","article-title":"Protein-Protein Association in Polymer Solutions: From Dilute to Semidilute to Concentrated","volume":"92","author":"Kozer","year":"2007","journal-title":"Biophysical J."},{"key":"B32","doi-asserted-by":"publisher","first-page":"116","DOI":"10.1016\/j.ymeth.2014.10.034","article-title":"Single-experiment Displacement Assay for Quantifying High-Affinity Binding by Isothermal Titration Calorimetry","volume":"76","author":"Krainer","year":"2015","journal-title":"Methods"},{"key":"B33","doi-asserted-by":"publisher","first-page":"3807","DOI":"10.1016\/j.febslet.2015.11.015","article-title":"Unusual Effects of Crowders on Heme Retention in Myoglobin","volume":"589","author":"Kundu","year":"2015","journal-title":"FEBS Lett."},{"key":"B34","doi-asserted-by":"publisher","first-page":"791","DOI":"10.1016\/s1074-5521(96)90063-0","article-title":"Sensing the Heat: The Application of Isothermal Titration Calorimetry to Thermodynamic Studies of Biomolecular Interactions","volume":"3","author":"Ladbury","year":"1996","journal-title":"Chem. Biol."},{"key":"B35","doi-asserted-by":"publisher","first-page":"253","DOI":"10.1042\/bj0890253","article-title":"The Interaction between Polysaccharides and Other Macromolecules. 5. The Solubility of Proteins in the Presence of Dextran","volume":"89","author":"Laurent","year":"1963","journal-title":"Biochem. J."},{"key":"B36","doi-asserted-by":"publisher","first-page":"1590","DOI":"10.1529\/biophysj.108.133025","article-title":"Molecular Dynamics Studies of Polyethylene Oxide and Polyethylene Glycol: Hydrodynamic Radius and Shape Anisotropy","volume":"95","author":"Lee","year":"2008","journal-title":"Biophysical J."},{"key":"B37","doi-asserted-by":"publisher","first-page":"e74053","DOI":"10.1371\/journal.pone.0074053","article-title":"Distal Effect of Amino Acid Substitutions in CYP2C9 Polymorphic Variants Causes Differences in Interatomic Interactions against (S)-Warfarin","volume":"8","author":"Lertkiatmongkol","year":"2013","journal-title":"PLoS One"},{"key":"B38","doi-asserted-by":"publisher","first-page":"7062","DOI":"10.1021\/jp100188s","article-title":"Hemoglobin\u2212Silver Interaction and Bioconjugate Formation: A Spectroscopic Study","volume":"114","author":"Mahato","year":"2010","journal-title":"J. Phys. Chem. B."},{"key":"B39","doi-asserted-by":"publisher","first-page":"570","DOI":"10.1042\/bj0710570","article-title":"Appendix-Spectrum of Horse-Heart Cytochrome C","volume":"71","author":"Margoliash","year":"1959","journal-title":"Biochem. J."},{"key":"B40","doi-asserted-by":"publisher","first-page":"127","DOI":"10.1016\/s0076-6879(98)95038-8","article-title":"[7] Molecular Crowding: Analysis of Effects of High Concentrations of Inert Cosolutes on Biochemical Equilibria and Rates in Terms of Volume Exclusion","volume":"295","author":"Minton","year":"1998","journal-title":"Methods Enzymol."},{"key":"B41","doi-asserted-by":"publisher","first-page":"2093","DOI":"10.1002\/bip.1981.360201006","article-title":"Excluded Volume as a Determinant of Macromolecular Structure and Reactivity","volume":"20","author":"Minton","year":"1981","journal-title":"Biopolymers"},{"key":"B42","doi-asserted-by":"publisher","first-page":"10577","DOI":"10.1074\/jbc.r100005200","article-title":"The Influence of Macromolecular Crowding and Macromolecular Confinement on Biochemical Reactions in Physiological media","volume":"276","author":"Minton","year":"2001","journal-title":"J. Biol. Chem."},{"key":"B43","doi-asserted-by":"publisher","first-page":"278","DOI":"10.1016\/j.saa.2018.03.026","article-title":"A Study on the Interaction of Horse Heart Cytochrome C with Some Conventional and Ionic Liquid Surfactants Probed by Ultraviolet-Visible and Fluorescence Spectroscopic Techniques","volume":"198","author":"Mondal","year":"2018","journal-title":"Spectrochimica Acta A: Mol. Biomol. Spectrosc."},{"key":"B44","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1023\/a:1023011609931","article-title":"Electrochemical Evidence for the Molten Globule States of Cytochrome C Induced by N-Alkyl Sulfates at Low Concentrations","volume":"22","author":"Moosavi-Movahedi","year":"2003","journal-title":"J. Protein Chem."},{"key":"B45","doi-asserted-by":"publisher","first-page":"1290","DOI":"10.1021\/bi00731a008","article-title":"Interaction of an Apolipoprotein (apoLP-Alanine) with Phosphatidylcholine","volume":"12","author":"Morrisett","year":"1973","journal-title":"Biochemistry"},{"key":"B46","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1016\/s1570-9639(02)00548-4","article-title":"Equilibrium Studies of the Effect of Difference in Sequence Homology on the Mechanism of Denaturation of Bovine and Horse Cytochromes-C","volume":"1646","author":"Moza","year":"2003","journal-title":"Biochim. Biophys. Acta (Bba) - Proteins Proteomics"},{"key":"B47","doi-asserted-by":"publisher","first-page":"4695","DOI":"10.1021\/bi052357r","article-title":"A Unique Molten Globule State Occurs during Unfolding of Cytochrome C by LiClO4 Near Physiological pH and Temperature: Structural and Thermodynamic Characterization","volume":"45","author":"Moza","year":"2006","journal-title":"Biochemistry"},{"key":"B48","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1007\/bf01025077","article-title":"Methionine-oxidized Horse Heart Cytochromec. III. Ascorbate Reduction and the Methionine-80-Sulfur-Iron Linkage","volume":"8","author":"Myer","year":"1989","journal-title":"J. Protein Chem."},{"key":"B49","doi-asserted-by":"publisher","first-page":"490","DOI":"10.3390\/biom10030490","article-title":"Interactions under Crowding Milieu: Chemical-Induced Denaturation of Myoglobin Is Determined by the Extent of Heme Dissociation on Interaction with Crowders","volume":"10","author":"Nasreen","year":"2020","journal-title":"Biomolecules"},{"key":"B50","doi-asserted-by":"publisher","first-page":"130","DOI":"10.1016\/j.ijbiomac.2017.08.014","article-title":"Macromolecular Crowding Induces Molten Globule State in the Native Myoglobin at Physiological pH","volume":"106","author":"Nasreen","year":"2018","journal-title":"Int. J. Biol. Macromolecules"},{"key":"B51","doi-asserted-by":"publisher","first-page":"215","DOI":"10.1385\/1-59259-912-5:215","article-title":"Probing Heme Protein-Ligand Interactions by UV\/visible Absorption Spectroscopy","volume":"305","author":"Nienhaus","year":"2005","journal-title":"Methods Mol. Biol."},{"key":"B52","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1016\/s0076-6879(72)26005-0","article-title":"[3] the Preparation of Guanidine Hydrochloride","volume":"26","author":"Nozaki","year":"1972","journal-title":"Methods Enzymol."},{"key":"B53","doi-asserted-by":"publisher","first-page":"192","DOI":"10.1016\/j.ijbiomac.2020.01.119","article-title":"Formation of Molten Globule State in Horse Heart Cytochrome C under Physiological Conditions: Importance of Soft Interactions and Spectroscopic Approach in Crowded Milieu","volume":"148","author":"Parray","year":"","journal-title":"Int. J. Biol. Macromolecules"},{"key":"B54","doi-asserted-by":"publisher","first-page":"13840","DOI":"10.1021\/acsomega.0c01185","article-title":"Effects of Ethylene Glycol on the Structure and Stability of Myoglobin Using Spectroscopic, Interaction, and In Silico Approaches: Monomer Is Different from Those of its Polymers","volume":"5","author":"Parray","year":"","journal-title":"ACS Omega"},{"key":"B55","doi-asserted-by":"publisher","first-page":"118788","DOI":"10.1016\/j.saa.2020.118788","article-title":"Conformational Changes in Cytochrome C Directed by Ethylene Glycol Accompanying Complex Formation: Protein-Solvent Preferential Interaction Or\/and Kosmotropic Effect","volume":"242","author":"Parray","year":"","journal-title":"Spectrochimica Acta Part A: Mol. Biomol. Spectrosc."},{"key":"B56","doi-asserted-by":"publisher","first-page":"106316","DOI":"10.1016\/j.polymertesting.2019.106316","article-title":"Amphiphilic Nature of Polyethylene Glycols and Their Role in Medical Research","volume":"82","author":"Parray","year":"","journal-title":"Polym. Test."},{"key":"B57","doi-asserted-by":"publisher","first-page":"6475","DOI":"10.1038\/s41598-021-85792-4","article-title":"Interaction of Polyethylene Glycol with Cytochrome C Investigated via In Vitro and In Silico Approaches","volume":"11","author":"Parray","year":"","journal-title":"Sci. Rep."},{"key":"B58","doi-asserted-by":"publisher","first-page":"2807","DOI":"10.3390\/molecules26092807","article-title":"Structural Refolding and Thermal Stability of Myoglobin in the Presence of Mixture of Crowders: Importance of Various Interactions for Protein Stabilization in Crowded Conditions","volume":"26","author":"Parray","year":"","journal-title":"Molecules"},{"key":"B59","doi-asserted-by":"publisher","first-page":"117502","DOI":"10.1016\/j.molliq.2021.117502","article-title":"Characterization of Different Intermediate States in Myoglobin Induced by Polyethylene Glycol: A Process of Spontaneous Molecular Self-Organization Foresees the Energy Landscape Theory via In Vitro and In Silico Approaches","volume":"342","author":"Parray","year":"","journal-title":"J. Mol. Liquids"},{"key":"B60","doi-asserted-by":"publisher","first-page":"1288","DOI":"10.1016\/j.ijbiomac.2018.12.170","article-title":"First Evidence of Formation of Pre-Molten Globule State in Myoglobin: A Macromolecular Crowding Approach towards Protein Folding In Vivo","volume":"126","author":"Parray","year":"2019","journal-title":"Int. J. Biol. Macromolecules"},{"key":"B61","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1016\/j.ijbiomac.2017.02.084","article-title":"Characterization of Intermediate State of Myoglobin in the Presence of PEG 10 under Physiological Conditions","volume":"99","author":"Parray","year":"2017","journal-title":"Int. J. Biol. Macromolecules"},{"key":"B62","doi-asserted-by":"publisher","first-page":"389","DOI":"10.1016\/j.jsps.2018.12.008","article-title":"Plant-derived Antiviral Drugs as Novel Hepatitis B Virus Inhibitors: Cell Culture and Molecular Docking Study","volume":"27","author":"Parvez","year":"2019","journal-title":"Saudi Pharm. J."},{"key":"B63","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1080\/09687680110103631","article-title":"The Hydrophobic Moment and its Use in the Classification of Amphiphilic Structures (Review)","volume":"19","author":"Phoenix","year":"2002","journal-title":"Mol. Membr. Biol."},{"key":"B64","first-page":"2724","article-title":"Elimination of the Negative Soret Cotton Effect of Cytochrome C by Replacement of the Invariant Phenylalanine Using Site-Directed Mutagenesis","volume-title":"J. Am. Chem. Soc.","author":"Pielak","year":"1986"},{"key":"B65","doi-asserted-by":"publisher","first-page":"831","DOI":"10.1038\/nsmb1132","article-title":"Dynamically Driven Protein Allostery","volume":"13","author":"Popovych","year":"2006","journal-title":"Nat. Struct. Mol. Biol."},{"key":"B66","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1016\/j.ijbiomac.2012.04.021","article-title":"Effect of Polyethylene Glycols on the Alkaline-Induced Molten Globule Intermediate of Bovine Serum Albumin","volume":"51","author":"Qu","year":"2012","journal-title":"Int. J. Biol. Macromolecules"},{"key":"B67","doi-asserted-by":"publisher","first-page":"1684","DOI":"10.1021\/bi0271042","article-title":"Conformational and Thermodynamic Characterization of the Molten Globule State Occurring During Unfolding of Cytochromes-C by Weak Salt Denaturants","volume":"42","author":"Qureshi","year":"2003","journal-title":"Biochemistry"},{"key":"B68","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1016\/j.bbamem.2013.05.023","article-title":"Isothermal Titration Calorimetry of Membrane Proteins - Progress and Challenges","volume":"1838","author":"Rajarathnam","year":"2014","journal-title":"Biochim. Biophys. Acta (Bba) - Biomembranes"},{"key":"B69","doi-asserted-by":"publisher","first-page":"561","DOI":"10.1016\/j.bbrc.2010.01.067","article-title":"Molecular Mechanism of Polyethylene Glycol Mediated Stabilization of Protein","volume":"392","author":"Rawat","year":"2010","journal-title":"Biochem. Biophysical Res. Commun."},{"key":"B70","doi-asserted-by":"publisher","first-page":"466","DOI":"10.1016\/0076-6879(95)62038-9","article-title":"[36] Using Macromolecular Crowding Agents to Identify Weak Interactions within DNA Replication Complexes","volume":"262","author":"Reddy","year":"1995","journal-title":"Methods Enzymol."},{"key":"B71","doi-asserted-by":"publisher","first-page":"9379","DOI":"10.1021\/bi990355z","article-title":"Direct Observation of the Self-Association of Dilute Proteins in the Presence of Inert Macromolecules at High Concentration via Tracer Sedimentation Equilibrium: Theory, Experiment, and Biological Significance","volume":"38","author":"Rivas","year":"1999","journal-title":"Biochemistry"},{"key":"B72","doi-asserted-by":"publisher","first-page":"211","DOI":"10.1016\/s0162-0134(97)00100-1","article-title":"The Soret Circular Dichroism Spectrum as a Probe for the Heme Fe(III)-Met(80) Axial Bond in Horse Cytochrome C","volume":"68","author":"Santucci","year":"1997","journal-title":"J. Inorg. Biochem."},{"key":"B73","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1007\/s12551-013-0104-4","article-title":"Soft Interactions and Crowding","volume":"5","author":"Sarkar","year":"","journal-title":"Biophys. Rev."},{"key":"B74","doi-asserted-by":"publisher","first-page":"19342","DOI":"10.1073\/pnas.1312678110","article-title":"Impact of Reconstituted Cytosol on Protein Stability","volume":"110","author":"Sarkar","year":"","journal-title":"Proc. Natl. Acad. Sci."},{"key":"B75","doi-asserted-by":"publisher","first-page":"1227","DOI":"10.1016\/s0022-2836(02)01443-2","article-title":"Effect of Dextran on Protein Stability and Conformation Attributed to Macromolecular Crowding","volume":"326","author":"Sasahara","year":"2003","journal-title":"J. Mol. Biol."},{"key":"B76","doi-asserted-by":"publisher","first-page":"477","DOI":"10.1021\/ja9529448","article-title":"Oxidation State-Induced Change of Iron Ligand in the Phenylalanine-82 to Histidine Mutant of Yeast Iso-1-Cytochrome C","volume":"118","author":"Schejter","year":"1996","journal-title":"J. Am. Chem. Soc."},{"key":"B77","doi-asserted-by":"publisher","first-page":"417","DOI":"10.1007\/s10822-010-9352-6","article-title":"Ligand Docking and Binding Site Analysis with PyMOL and Autodock\/Vina","volume":"24","author":"Seeliger","year":"2010","journal-title":"J. Comput. Aided Mol. Des."},{"key":"B78","doi-asserted-by":"publisher","first-page":"927","DOI":"10.1007\/s12010-019-02972-9","article-title":"Mixture of Macromolecular Crowding Agents Has a Non-Additive Effect on the Stability of Proteins","volume":"188","author":"Shahid","year":"2019","journal-title":"Appl. Biochem. Biotechnol."},{"key":"B79","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1016\/j.abb.2015.08.015","article-title":"Relationship Between Protein Stability and Functional Activity in the Presence of Macromolecular Crowding Agents Alone and in Mixture: An Insight into Stability-Activity Trade-Off","volume":"584","author":"Shahid","year":"2015","journal-title":"Arch. Biochem. Biophys."},{"key":"B80","doi-asserted-by":"publisher","first-page":"178","DOI":"10.1016\/j.bbagen.2016.11.014","article-title":"Size-Dependent Studies of Macromolecular Crowding on the Thermodynamic Stability, Structure and Functional Activity of Proteins: In Vitro and In Silico Approaches","volume":"1861","author":"Shahid","year":"2017","journal-title":"Biochim. Biophys. Acta (Bba) - Gen. Subjects"},{"key":"B81","doi-asserted-by":"publisher","first-page":"7990","DOI":"10.1073\/pnas.1505396112","article-title":"Analysis of the Size Dependence of Macromolecular Crowding Shows that Smaller Is Better","volume":"112","author":"Sharp","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"B82","doi-asserted-by":"crossref","first-page":"8165","DOI":"10.1021\/jp800311c","article-title":"Flexibility of Human Cytochromes P450: Molecular Dynamics Reveals Differences Between CYPs 3A4, 2C9, and 2A6, Which Correlate with Their Substrate Preferences","volume":"112","author":"Skopal\u00edk","year":"2008","journal-title":"J. Phys. Chem. B"},{"key":"B83","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1007\/s12551-016-0218-6","article-title":"Dynamic Light Scattering: A Practical Guide and Applications in Biomedical Sciences","volume":"8","author":"Stetefeld","year":"2016","journal-title":"Biophys. Rev."},{"key":"B84","doi-asserted-by":"publisher","first-page":"125","DOI":"10.1110\/ps.03288104","article-title":"Protein Folding by the Effects of Macromolecular Crowding","volume":"13","author":"Tokuriki","year":"2004","journal-title":"Protein Sci."},{"key":"B85","doi-asserted-by":"crossref","first-page":"255","DOI":"10.4149\/gpb_2010_03_255","article-title":"Rate of Oxidative Modification of Cytochrome C by Hydrogen Peroxide Is Modulated by Hofmeister Anions","volume":"29","author":"Tom\u00e1\u0161kov\u00e1","year":"2010","journal-title":"Gen. Physiol. Biophys."},{"key":"B86","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1016\/j.jchromb.2003.10.060","article-title":"Relationship Between the Protein Surface Hydrophobicity and its Partitioning Behaviour in Aqueous Two-Phase Systems of Polyethyleneglycol-Dextran","volume":"799","author":"Tubio","year":"2004","journal-title":"J. Chromatogr. B"},{"key":"B87","doi-asserted-by":"publisher","first-page":"273","DOI":"10.1016\/j.ab.2004.06.010","article-title":"Determination of the Refractive Index Increment (Dn\/dc) of Molecule and Macromolecule Solutions by Surface Plasmon Resonance","volume":"333","author":"Tumolo","year":"2004","journal-title":"Anal. Biochem."},{"key":"B88","doi-asserted-by":"publisher","first-page":"6927","DOI":"10.1093\/emboj\/18.24.6927","article-title":"Effects of Macromolecular Crowding on Protein Folding and Aggregation","volume":"18","author":"Van Den Berg","year":"1999","journal-title":"EMBO J."},{"key":"B89","doi-asserted-by":"publisher","first-page":"3870","DOI":"10.1093\/emboj\/19.15.3870","article-title":"Macromolecular Crowding Perturbs Protein Refolding Kinetics: Implications for Folding Inside the Cell","volume":"19","author":"Van Den Berg","year":"2000","journal-title":"EMBO J."},{"key":"B90","doi-asserted-by":"publisher","first-page":"F605","DOI":"10.1152\/ajprenal.00171.2004","article-title":"Ficoll and Dextran vs. Globular Proteins as Probes for Testing Glomerular Permselectivity: Effects of Molecular Size, Shape, Charge, and Deformability","volume":"288","author":"Venturoli","year":"2005","journal-title":"Am. J. Physiology-Renal Physiol."},{"key":"B91","doi-asserted-by":"publisher","first-page":"127","DOI":"10.1093\/protein\/8.2.127","article-title":"LIGPLOT: A Program to Generate Schematic Diagrams of Protein-Ligand Interactions","volume":"8","author":"Wallace","year":"1995","journal-title":"Protein Eng. Des. Sel"},{"key":"B92","doi-asserted-by":"publisher","first-page":"383","DOI":"10.1016\/b978-0-12-800046-5.00011-4","article-title":"Crowding, Diffusion, and Biochemical Reactions","volume":"307","author":"Weiss","year":"2014","journal-title":"Int. Rev. Cell Mol. Biol."},{"key":"B93","doi-asserted-by":"publisher","first-page":"186","DOI":"10.1016\/j.bpc.2005.08.001","article-title":"Molecular Crowding Effects of Linear Polymers in Protein Solutions","volume":"119","author":"Winzor","year":"2006","journal-title":"Biophysical Chem."},{"key":"B94","doi-asserted-by":"publisher","first-page":"27913","DOI":"10.1039\/c7ra02838e","article-title":"Interaction of Human Serum Albumin with Uremic Toxins: A Thermodynamic Study","volume":"7","author":"Yu","year":"2017","journal-title":"RSC Adv."},{"key":"B95","doi-asserted-by":"publisher","first-page":"229","DOI":"10.1007\/s00018-013-1341-1","article-title":"The Role of Key Residues in Structure, Function, and Stability of Cytochrome-C","volume":"71","author":"Zaidi","year":"2014","journal-title":"Cell. Mol. Life Sci."},{"key":"B96","doi-asserted-by":"publisher","first-page":"703","DOI":"10.1093\/abbs\/gms052","article-title":"Effects of Macromolecular Crowding on the Structural Stability of Human \u03b1-Lactalbumin","volume":"44","author":"Zhang","year":"2012","journal-title":"Acta Biochim. Biophys. Sin (Shanghai)"},{"key":"B97","doi-asserted-by":"publisher","first-page":"55109","DOI":"10.1074\/jbc.m409086200","article-title":"Mixed Macromolecular Crowding Accelerates the Oxidative Refolding of Reduced, Denatured Lysozyme","volume":"279","author":"Zhou","year":"2004","journal-title":"J. Biol. Chem."},{"key":"B98","doi-asserted-by":"publisher","first-page":"375","DOI":"10.1146\/annurev.biophys.37.032807.125817","article-title":"Macromolecular Crowding and Confinement: Biochemical, Biophysical, and Potential Physiological Consequences","volume":"37","author":"Zhou","year":"2008","journal-title":"Annu. Rev. Biophys."},{"key":"B99","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1146\/annurev.bb.22.060193.000331","article-title":"Macromolecular Crowding: Biochemical, Biophysical, and Physiological Consequences","volume":"22","author":"Zimmerman","year":"1993","journal-title":"Annu. Rev. Biophys. Biomol. Struct."}],"container-title":["Frontiers in Molecular Biosciences"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmolb.2022.849683\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,5,26]],"date-time":"2022-05-26T17:19:49Z","timestamp":1653585589000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmolb.2022.849683\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,5,25]]},"references-count":99,"alternative-id":["10.3389\/fmolb.2022.849683"],"URL":"https:\/\/doi.org\/10.3389\/fmolb.2022.849683","relation":{},"ISSN":["2296-889X"],"issn-type":[{"value":"2296-889X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,5,25]]},"article-number":"849683"}}