{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,29]],"date-time":"2026-03-29T12:28:43Z","timestamp":1774787323106,"version":"3.50.1"},"reference-count":36,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2014,4,22]],"date-time":"2014-04-22T00:00:00Z","timestamp":1398124800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Chaperonin GroEL is a complex oligomeric heat shock protein (Hsp60) assisting the correct folding and assembly of other proteins in the cell. An intriguing question is how GroEL folds itself. According to the literature, GroEL reassembly is dependent on chaperonin ligands and solvent composition. Here we demonstrate dependence of GroEL reassembly efficiency on concentrations of the essential factors (Mg2+, ADP, ATP, GroES, ammonium sulfate, NaCl and glycerol). Besides, kinetics of GroEL oligomerization in various conditions was monitored by the light scattering technique and proved to be two-exponential,  which suggested accumulation of a certain oligomeric intermediate. This intermediate was resolved as a heptamer by nondenaturing blue electrophoresis of GroEL monomers during their assembly in the presence of both Mg-ATP and co-chaperonin GroES. Presumably, this intermediate heptamer plays a key role in formation of the GroEL tetradecameric particle. The role of co-chaperonin GroES (Hsp10) in GroEL assembly is also discussed.<\/jats:p>","DOI":"10.3390\/biom4020458","type":"journal-article","created":{"date-parts":[[2014,4,22]],"date-time":"2014-04-22T12:18:37Z","timestamp":1398169117000},"page":"458-473","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Chaperonin GroEL Reassembly: An Effect of Protein Ligands and Solvent Composition"],"prefix":"10.3390","volume":"4","author":[{"given":"Nataliya","family":"Ryabova","sequence":"first","affiliation":[{"name":"Institute of Protein Research, Russian Academy of Sciences, Institutskaya Street 4, Pushchino 142290, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Victor","family":"Marchenkov","sequence":"additional","affiliation":[{"name":"Institute of Protein Research, Russian Academy of Sciences, Institutskaya Street 4, Pushchino 142290, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nina","family":"Kotova","sequence":"additional","affiliation":[{"name":"Institute of Protein Research, Russian Academy of Sciences, Institutskaya Street 4, Pushchino 142290, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gennady","family":"Semisotnov","sequence":"additional","affiliation":[{"name":"Institute of Protein Research, Russian Academy of Sciences, Institutskaya Street 4, Pushchino 142290, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2014,4,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1126\/science.181.4096.223","article-title":"Principles of govern the folding of protein chains","volume":"181","author":"Anfinsen","year":"1973","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1042","DOI":"10.1126\/science.1219021","article-title":"The Protein-folding problem, 50 years on","volume":"338","author":"Dill","year":"2012","journal-title":"Science"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1146\/annurev.biochem.70.1.603","article-title":"Folding of newly translated proteins in vivo: The chaperones","volume":"70","author":"Frydman","year":"2001","journal-title":"Annu. Rev. Biochem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"324","DOI":"10.1038\/nature10317","article-title":"Molecular chaperones in protein folding and proteostasis","volume":"475","author":"Hartl","year":"2011","journal-title":"Nature"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1038\/328378a0","article-title":"Proteins as molecular chaperones","volume":"328","author":"Ellis","year":"1987","journal-title":"Nature"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1016\/j.tibs.2006.05.001","article-title":"Molecular chaperones: Assisting assembly in addition to folding","volume":"31","author":"Ellis","year":"2006","journal-title":"Trends Biochem. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1038\/333330a0","article-title":"Homologous plant and bacterial proteins chaperone oligomeric protein assembly","volume":"333","author":"Hemmingsen","year":"1988","journal-title":"Nature"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1852","DOI":"10.1126\/science.1068408","article-title":"Molecular chaperones in the cytosol: From nansent chain to folded protein","volume":"295","author":"Hartl","year":"2002","journal-title":"Science"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Gething, M.-J. (1997). Guidebook to Molecular Chaperones and Protein-Folding Catalysts, Oxford University Press.","DOI":"10.1093\/oso\/9780198599494.001.0001"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1016\/0968-0004(89)90168-0","article-title":"Molecular chaperones: Proteins essential for the biogenesis of some macromolecular structures","volume":"14","author":"Ellis","year":"1989","journal-title":"Trends Biochem. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1146\/annurev.cellbio.23.090506.123555","article-title":"Two families of chaperonin: Physiology and mechanism","volume":"23","author":"Horwich","year":"2007","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1016\/j.tibs.2011.05.003","article-title":"Chaperonins: Two rings for folding","volume":"36","author":"Yebenes","year":"2011","journal-title":"Trends Biochem. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"578","DOI":"10.1038\/371578a0","article-title":"The crystal structure of the bacterial chaperonin GroEL at 2.8 A","volume":"371","author":"Braig","year":"1994","journal-title":"Nature"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1038\/379037a0","article-title":"The crystal structure of the GroES co-chaperonin at 2.8 A resolution","volume":"379","author":"Hunt","year":"1996","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"741","DOI":"10.1038\/41944","article-title":"The crystal structure of the asymmetric GroEL-GroES-(ADP)7 chaperonin complex","volume":"388","author":"Xu","year":"1997","journal-title":"Nature"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1038\/348339a0","article-title":"(Mg-ATP)-dependent self-assembly of molecular chaperone GroEL","volume":"348","author":"Lissin","year":"1990","journal-title":"Nature"},{"key":"ref_17","first-page":"314","article-title":"The monomeric form of the molecular chaperone GroEL: Structure, stability and oligomerization","volume":"25","author":"Surin","year":"1999","journal-title":"Russ. J. Bioorg. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/0014-5793(93)81528-8","article-title":"Stabilization of a compact conformation of monomeric GroEL at low temperature by adenine nucleotides","volume":"324","author":"Lissin","year":"1993","journal-title":"FEBS Lett."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/0014-5793(95)00151-X","article-title":"In vitro dissociation and self-assembly of three chaperonin 60s: The role of ATP","volume":"361","author":"Lissin","year":"1995","journal-title":"FEBS Lett."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"672","DOI":"10.1110\/ps.0233603","article-title":"Denaturation and reassembly of chaperonin GroEL studied by solution X-ray scattering","volume":"12","author":"Arai","year":"2003","journal-title":"Protein Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2213","DOI":"10.1074\/jbc.270.38.22113","article-title":"Refolding and reassembly of active chaperonin GroEL after denaturation","volume":"270","author":"Ybarra","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"22962","DOI":"10.1074\/jbc.270.39.22962","article-title":"Inactive GroEL monomers can be isolated and reassembled to functional tetradecamers that contain few bound peptides","volume":"270","author":"Ybarra","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1016\/0167-4838(94)00231-5","article-title":"Tetradecameric chaperonin 60 can be assembled in vitro from monomers in a process that is ATP independent","volume":"1247","author":"Mendoza","year":"1995","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1016\/0022-2836(79)90279-1","article-title":"Electrophoresis analysis of the unfolding of proteins by urea","volume":"129","author":"Greighton","year":"1979","journal-title":"J. Mol. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0003-2697(84)90761-9","article-title":"Gel electrophoresis in studies of protein conformation and folding","volume":"138","author":"Goldenberg","year":"1984","journal-title":"Anal. Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"13928","DOI":"10.1021\/bi00042a026","article-title":"Residual structure in urea-denatured chaperonin GroEL","volume":"34","author":"Gorovits","year":"1995","journal-title":"Biochemistry"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2447","DOI":"10.1016\/S0021-9258(17)41966-1","article-title":"Alternation of the quaternary structure of Cpn60 modulates chaperonin-assisted folding","volume":"269","author":"Mendosa","year":"1994","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/S1097-2765(00)80117-3","article-title":"A single ring is sufficient for productive chaperonin-mediated folding in vivo","volume":"2","author":"Nielsen","year":"1998","journal-title":"Mol. Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"897","DOI":"10.1006\/jmbi.2000.4278","article-title":"From minichaperone to GroEL: Properties of an active single-ring mutant of GroEL","volume":"304","author":"Chatellier","year":"2000","journal-title":"J. Mol. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1016\/0092-8674(95)90098-5","article-title":"Mechanism of GroEL action: Productive release of polypeptide from a sequestered position under GroES","volume":"83","author":"Weissman","year":"1995","journal-title":"Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1016\/0003-2697(91)90094-A","article-title":"Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form","volume":"199","author":"Schagger","year":"1991","journal-title":"Anal. Biochem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/S0076-6879(98)90010-6","article-title":"Purification of GroEL with low fluorescence background","volume":"290","author":"Clark","year":"1998","journal-title":"Methods Enzymol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1038\/371614a0","article-title":"Residues in chaperonin GroEL required for polypeptide binding and release","volume":"371","author":"Fenton","year":"1994","journal-title":"Nature"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1038\/227680a0","article-title":"Cleavage of structural proteins during the assembly of the head of bacteriophage T4","volume":"227","author":"Laemmli","year":"1970","journal-title":"Nature"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2066","DOI":"10.3390\/ijms10052066","article-title":"GroEL-assisted protein folding: Does it occur within the chaperonin inner cavity","volume":"10","author":"Marchenkov","year":"2009","journal-title":"Int. J. Mol. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1405","DOI":"10.1134\/S0006297913130038","article-title":"Molecular chaperone GroEL\/ES: Unfolding and refolding processes","volume":"78","author":"Ryabova","year":"2013","journal-title":"Biochem. Mosc."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/4\/2\/458\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:10:33Z","timestamp":1760217033000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/4\/2\/458"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,4,22]]},"references-count":36,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2014,6]]}},"alternative-id":["biom4020458"],"URL":"https:\/\/doi.org\/10.3390\/biom4020458","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,4,22]]}}}