{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T17:19:16Z","timestamp":1788283156436,"version":"build-2803163510"},"reference-count":30,"publisher":"American Chemical Society (ACS)","issue":"38","license":[{"start":{"date-parts":[[2015,9,3]],"date-time":"2015-09-03T00:00:00Z","timestamp":1441238400000},"content-version":"vor","delay-in-days":365,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"funder":[{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["R21-EY021595"],"award-info":[{"award-number":["R21-EY021595"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["P20GM10341"],"award-info":[{"award-number":["P20GM10341"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2014,9,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Protective antigen (PA) mediates entry of edema factor (EF) and lethal factor (LF) into the cytoplasmic space of the cells through the formation of a membrane-spanning pore. To do this, PA must initially bind to a host cellular receptor. Recent mass spectrometry analysis of PA using histidine hydrogen\u2013deuterium exchange (His-HDX) has shown that binding of the von Willebrand factor A (vWA) domain of the receptor capillary morphogenesis protein-2 (CMG2) lowers the exchange rates of the imidazole C2 hydrogen of several histidines, suggesting that receptor binding decreases the structural flexibility of PA. Here, using His-HDX and fluorescence as a function of denaturant, and protease susceptibility, we show that binding of the vWA domain of CMG2 largely increases the stability of PA and the effect reaches up to 70 \u00c5 from the receptor binding interface. We also show that the pKa values and HDX rates of histidines located in separate domains change upon receptor binding. These results indicate that when one end of the protein is anchored, the structure of PA is tightened, noncovalent interactions are strengthened, and the global stability of the protein increases. These findings suggest that CMG2 may be used to stabilize PA in future anthrax vaccines.<\/jats:p>","DOI":"10.1021\/bi500718g","type":"journal-article","created":{"date-parts":[[2014,9,4]],"date-time":"2014-09-04T11:30:25Z","timestamp":1409830225000},"page":"6084-6091","source":"Crossref","is-referenced-by-count":9,"title":["Long-Range Stabilization\nof Anthrax Protective Antigen\nupon Binding to CMG2"],"prefix":"10.1021","volume":"53","author":[{"given":"Vennela","family":"Mullangi","sequence":"first","affiliation":[{"name":"Case Western Reserve University , , , ,","place":["Cleveland, Ohio, United States, 44106"]},{"name":"Cleveland State University , , , ,","place":["Cleveland, Ohio, United States, 44114"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sireesha","family":"Mamillapalli","sequence":"additional","affiliation":[{"name":"Wichita State University , , , ,","place":["Wichita, Kansas, United States, 67260"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David J.","family":"Anderson","sequence":"additional","affiliation":[{"name":"Cleveland State University , , , ,","place":["Cleveland, Ohio, United States, 44114"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James G.","family":"Bann","sequence":"additional","affiliation":[{"name":"Wichita State University , , , ,","place":["Wichita, Kansas, United States, 67260"]},{"name":"University of Kansas School of Medicine-Wichita , , ,","place":["Wichita, Kansas, United States, 67214"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masaru","family":"Miyagi","sequence":"additional","affiliation":[{"name":"Case Western Reserve University , , , ,","place":["Cleveland, Ohio, United States, 44106"]},{"name":"Case Western Reserve University , , , ,","place":["Cleveland, Ohio, United States, 44106"]},{"name":"Case Western Reserve University , , , ,","place":["Cleveland, Ohio, United States, 44106"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2014,9,3]]},"reference":[{"key":"2026090112464681200_cit1","doi-asserted-by":"crossref","first-page":"10277","DOI":"10.1073\/pnas.89.21.10277","article-title":"Anthrax toxin protective\nantigen is activated by a cell surface protease with the sequence\nspecificity and catalytic properties of furin","volume":"89","author":"Klimpel","year":"1992","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026090112464681200_cit2","doi-asserted-by":"crossref","first-page":"13147","DOI":"10.1073\/pnas.0405405101","article-title":"Structure\nof heptameric protective antigen bound to an anthrax toxin receptor:\nA role for receptor in pH-dependent pore formation","volume":"101","author":"Lacy","year":"2004","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026090112464681200_cit3","doi-asserted-by":"crossref","first-page":"741","DOI":"10.1016\/j.jmb.2010.04.041","article-title":"Role of\nthe protective antigen octamer in the molecular mechanism of anthrax\nlethal toxin stabilization in plasma","volume":"399","author":"Kintzer","year":"2010","journal-title":"J. Mol.\nBiol."},{"key":"2026090112464681200_cit4","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1038\/nature02763","article-title":"Crystal\nstructure of a complex between anthrax toxin and its host cell receptor","volume":"430","author":"Santelli","year":"2004","journal-title":"Nature"},{"key":"2026090112464681200_cit5","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1038\/n35101999","article-title":"Identification\nof the cellular receptor for anthrax toxin","volume":"414","author":"Bradley","year":"2001","journal-title":"Nature"},{"key":"2026090112464681200_cit6","doi-asserted-by":"crossref","first-page":"1860","DOI":"10.1128\/IAI.67.4.1860-1865.1999","article-title":"Identification of\na receptor-binding region within domain 4 of the protective antigen\ncomponent of anthrax toxin","volume":"67","author":"Varughese","year":"1999","journal-title":"Infect. Immun."},{"key":"2026090112464681200_cit7","doi-asserted-by":"crossref","first-page":"23349","DOI":"10.1074\/jbc.M401292200","article-title":"Binding stoichiometry\nand kinetics of the interaction\nof a human anthrax toxin receptor, CMG2, with protective antigen","volume":"279","author":"Wigelsworth","year":"2004","journal-title":"J. Biol. Chem."},{"key":"2026090112464681200_cit8","doi-asserted-by":"crossref","first-page":"1467","DOI":"10.1002\/pro.2136","article-title":"pH effects on binding between the anthrax protective\nantigen and the host cellular receptor CMG2","volume":"21","author":"Rajapaksha","year":"2012","journal-title":"Protein Sci."},{"key":"2026090112464681200_cit9","doi-asserted-by":"crossref","first-page":"e329","DOI":"10.1371\/journal.pone.0000329","article-title":"Anthrax toxin receptor\n2 determinants that dictate\nthe pH threshold of toxin pore formation","volume":"2","author":"Scobie","year":"2007","journal-title":"PLoS\nOne"},{"key":"2026090112464681200_cit10","doi-asserted-by":"crossref","first-page":"13278","DOI":"10.1073\/pnas.0505865102","article-title":"Receptor-specific\nrequirements for anthrax toxin delivery\ninto cells","volume":"102","author":"Rainey","year":"2005","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"2026090112464681200_cit11","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1021\/bi401405s","article-title":"19F nuclear\nmagnetic resonance and crystallographic\nstudies of 5-fluorotryptophan-labeled anthrax protective antigen and\neffects of the receptor on stability","volume":"53","author":"Chadegani","year":"2014","journal-title":"Biochemistry"},{"key":"2026090112464681200_cit12","doi-asserted-by":"crossref","first-page":"6481","DOI":"10.1021\/ac8009643","article-title":"Determination of pKa values of individual\nhistidine residues in proteins using mass spectrometry","volume":"80","author":"Miyagi","year":"2008","journal-title":"Anal. Chem."},{"key":"2026090112464681200_cit13","doi-asserted-by":"crossref","first-page":"1653","DOI":"10.1021\/ac202927p","article-title":"Slow histidine H\/D exchange protocol for thermodynamic\nanalysis of protein folding and stability using mass spectrometry","volume":"84","author":"Tran","year":"2012","journal-title":"Anal. Chem."},{"key":"2026090112464681200_cit14","doi-asserted-by":"crossref","first-page":"6973","DOI":"10.1021\/bi100647z","article-title":"Evidence that histidine protonation\nof receptor-bound anthrax protective antigen is a trigger for pore\nformation","volume":"49","author":"Wimalasena","year":"2010","journal-title":"Biochemistry"},{"key":"2026090112464681200_cit15","doi-asserted-by":"crossref","first-page":"14928","DOI":"10.1021\/bi701763z","article-title":"Effect\nof 2-fluorohistidine labeling of the anthrax protective antigen on\nstability, pore formation, and translocation","volume":"46","author":"Wimalasena","year":"2007","journal-title":"Biochemistry"},{"key":"2026090112464681200_cit16","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1002\/pro.26","article-title":"Monitoring\nanthrax toxin receptor dissociation from\nthe protective antigen by NMR","volume":"18","author":"Rajapaksha","year":"2009","journal-title":"Protein Sci."},{"key":"2026090112464681200_cit17","doi-asserted-by":"crossref","first-page":"e17055","DOI":"10.1371\/journal.pone.0017055","article-title":"Histidine\nhydrogen-deuterium exchange mass spectrometry\nfor probing the microenvironment of histidine residues in dihydrofolate\nreductase","volume":"6","author":"Miyagi","year":"2011","journal-title":"PLoS One"},{"key":"2026090112464681200_cit18","doi-asserted-by":"crossref","first-page":"1438","DOI":"10.1016\/j.jmb.2007.02.077","article-title":"Comparison of proteolytic susceptibility\nin phosphoglycerate kinases from yeast and E. coli: Modulation of conformational ensembles without altering structure\nor stability","volume":"368","author":"Young","year":"2007","journal-title":"J. Mol. Biol."},{"key":"2026090112464681200_cit19","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1002\/(SICI)1096-987X(199802)19:3<319::AID-JCC6>3.0.CO;2-W","article-title":"Exact and efficient analytical calculation\nof the accessible surface\nareas and their gradients for macromolecules","volume":"19","author":"Fraczkiewicz","year":"1998","journal-title":"J. Comput. Chem."},{"key":"2026090112464681200_cit20","doi-asserted-by":"crossref","first-page":"10649","DOI":"10.1021\/bi990580g","article-title":"Atomic mutations at\nthe single tryptophan residue of human recombinant annexin V: Effects\non structure, stability, and activity","volume":"38","author":"Minks","year":"1999","journal-title":"Biochemistry"},{"key":"2026090112464681200_cit21","doi-asserted-by":"crossref","first-page":"572","DOI":"10.1016\/0003-9861(68)90179-3","article-title":"Comparison\nof the specificities of\nvarious neutral proteinases from microorganisms","volume":"123","author":"Morihara","year":"1968","journal-title":"Arch. Biochem. Biophys."},{"key":"2026090112464681200_cit22","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1038\/385833a0","article-title":"Crystal\nstructure of the anthrax toxin protective antigen","volume":"385","author":"Petosa","year":"1997","journal-title":"Nature"},{"key":"2026090112464681200_cit23","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1021\/jo00829a062","article-title":"The kinetics of deuteration of imidazole","volume":"35","author":"Vaughan","year":"1970","journal-title":"J. Org. Chem."},{"key":"2026090112464681200_cit24","doi-asserted-by":"crossref","first-page":"6139","DOI":"10.1021\/ja00799a063","article-title":"Hydrogen-deuterium exchange kinetics\nof the C-2 protons of imidazole and histidine compounds","volume":"95","author":"Bradbury","year":"1973","journal-title":"J. Am. Chem. Soc."},{"key":"2026090112464681200_cit25","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.jmb.2011.10.035","article-title":"Domain\nflexibility modulates the heterogeneous assembly mechanism of anthrax\ntoxin protective antigen","volume":"415","author":"Feld","year":"2012","journal-title":"J. Mol. Biol."},{"key":"2026090112464681200_cit26","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1111\/j.1600-065X.2010.00969.x","article-title":"Progress and\nnovel strategies in vaccine development and treatment of anthrax","volume":"239","author":"Chitlaru","year":"2011","journal-title":"Immunol. Rev."},{"key":"2026090112464681200_cit27","doi-asserted-by":"crossref","first-page":"5045","DOI":"10.1021\/bi049965b","article-title":"Conformational\ndynamics of the GdmHCl-induced molten globule state of creatine kinase\nmonitored by hydrogen exchange and mass spectrometry","volume":"43","author":"Mazon","year":"2004","journal-title":"Biochemistry"},{"key":"2026090112464681200_cit28","doi-asserted-by":"crossref","first-page":"1461","DOI":"10.1002\/rcm.1941","article-title":"Denaturant sensitive\nregions in creatine kinase identified by hydrogen\/deuterium exchange","volume":"19","author":"Mazon","year":"2005","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"2026090112464681200_cit29","doi-asserted-by":"crossref","first-page":"1224","DOI":"10.1039\/C0CS00113A","article-title":"Hydrogen exchange mass spectrometry for studying protein\nstructure and dynamics","volume":"40","author":"Konermann","year":"2011","journal-title":"Chem. Soc. Rev."},{"key":"2026090112464681200_cit30","doi-asserted-by":"crossref","first-page":"12439","DOI":"10.1021\/bi030192l","article-title":"The kinetics\nof side chain stabilization during protein folding","volume":"42","author":"Frieden","year":"2003","journal-title":"Biochemistry"}],"container-title":["Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/bi500718g","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/53\/38\/6084\/15694408\/bi500718g.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/53\/38\/6084\/15694408\/bi500718g.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T16:46:57Z","timestamp":1788281217000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/bichaw\/article\/53\/38\/6084\/1476846\/Long-Range-Stabilization-of-Anthrax-Protective"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,9,3]]},"references-count":30,"journal-issue":{"issue":"38","published-online":{"date-parts":[[2014,9,3]]},"published-print":{"date-parts":[[2014,9,30]]}},"URL":"https:\/\/doi.org\/10.1021\/bi500718g","relation":{},"ISSN":["0006-2960","1520-4995"],"issn-type":[{"value":"0006-2960","type":"print"},{"value":"1520-4995","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,9,3]]}}}