Contribution of domain interface residues to the stability of antibody CH3 domain homodimers

Biochemistry
W Dall'AcquaP Carter

Abstract

Dimers of CH3 domains from human IgG1 were used to study the effect of mutations constructed at a domain-domain interface upon domain dissociation and unfolding, "complex stability". Alanine replacement mutants were constructed on one side of the interface for each of the sixteen interdomain contact residues by using a single-chain CH3 dimer in which the carboxyl terminus of one domain was joined to the amino terminus of the second domain via a (G4S)4 linker. Single-chain variants were expressed in Escherichia coli grown in a fermentor and recovered in yields of 6-90 mg L-1 by immobilized metal affinity chromatography. Guanidine hydrochloride-induced denaturation was used to follow domain dissociation and unfolding. Surprisingly, the linker did not perturb the complex stability for either the wild type or two destabilizing mutants. The CH3 domain dissociation and unfolding energetics are dominated by six contact residues where corresponding alanine mutations each destabilize the complex by >2.0 kcal mol-1. Five of these residues (T366, L368, F405, Y407, and K409) form a patch at the center of the interface and are located on the two internal antiparallel beta-strands. These energetically key residues are surrounded by 10 residu...Continue Reading

References

Sep 20, 1992·Journal of Molecular Biology·A HorovitzA R Fersht
Jan 1, 1990·Annual Review of Biochemistry·D R DaviesS Sheriff
Jan 1, 1989·Annual Review of Biochemistry·T Alber
Jul 1, 1988·Journal of Chromatography·E Hochuli
Oct 5, 1988·Journal of Molecular Biology·M SchifferF J Stevens
Jan 1, 1985·Annual Review of Biochemistry·G K Ackers, F R Smith
Dec 5, 1985·Journal of Molecular Biology·C ChothiaM Karplus
Jan 1, 1972·Methods in Enzymology·T N Solie
Jan 6, 1981·Biochemistry·S W Provencher, J Glöckner
Dec 5, 1993·Journal of Molecular Biology·B C Cunningham, J A Wells
Jan 1, 1995·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·B C Braden, R J Poljak
Feb 1, 1993·Current Opinion in Immunology·R A Mariuzza, R J Poljak
Mar 1, 1993·Trends in Biochemical Sciences·D S Goodsell, A J Olson
Jan 9, 1996·Proceedings of the National Academy of Sciences of the United States of America·D R Davies, G H Cohen

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Citations

Dec 9, 2003·Journal of Molecular Biology·Stephen J DemarestGeneviève Hansen
Jun 2, 2011·Journal of the American Chemical Society·Theo RispensRob C Aalberse
Sep 11, 2013·Journal of the American Chemical Society·Morten BertzMatthias Rief
Feb 19, 2002·Immunology·Rob C Aalberse, Janine Schuurman
Oct 10, 2009·Proceedings of the National Academy of Sciences of the United States of America·Randall J BrezskiRobert E Jordan
Apr 21, 2012·The Journal of Biological Chemistry·Tianlei YingDimiter S Dimitrov
Jan 12, 2012·Protein Engineering, Design & Selection : PEDS·Ingunn B RasmussenInger Sandlie
Dec 26, 2012·Protein Engineering, Design & Selection : PEDS·Michael W TraxlmayrChristian Obinger
Nov 20, 2012·Applied and Environmental Microbiology·Jun Hyoung LeeSun Chang Kim
Dec 17, 2014·Biological & Pharmaceutical Bulletin·Yoichi IshiiKaori Wakamatsu
Dec 3, 2016·Journal of the American Society for Mass Spectrometry·Aneika C Leney, Albert J R Heck
Jan 11, 2017·MAbs·Ulrich Brinkmann, Roland E Kontermann
Sep 26, 2006·Journal of Pharmaceutical Sciences·Wei WangSandeep Nema
Apr 10, 2018·PloS One·Gordana Wozniak-KnoppFlorian Rüker
May 23, 2003·The Journal of Biological Chemistry·Marta del AlamoMauricio G Mateu
Mar 21, 2009·The Journal of Immunology : Official Journal of the American Association of Immunologists·Theo RispensRob C Aalberse
Aug 16, 2011·The Journal of Immunology : Official Journal of the American Association of Immunologists·Aran F LabrijnPaul W H I Parren
Nov 5, 2008·The Journal of Immunology : Official Journal of the American Association of Immunologists·Morten FlobakkInger Sandlie
Aug 2, 2016·PloS One·Lu ShanVaheh Oganesyan
Jun 7, 2017·Protein & Cell·Philip D BardwellJijie Gu
Sep 24, 2019·Antibodies·Qiong WangMichael J Betenbaugh
Feb 9, 2020·Scientific Reports·Youngsil SeoMyung-Hee Kwon
Jan 27, 2005·Journal of Cell Science·Oliver J HarrisonPeter J Kilshaw
May 26, 2021·MAbs·Monica L Fernández-QuinteroKlaus R Liedl

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