Modulation of Antibody Affinity by a Non-contact Residue

Overview

Journal Protein Sci

Specialty Biochemistry

Date 1993 Feb 1

PMID 8443598

Citations 7

Authors

J F Schillbach

R I Near

R E Bruccoleri

E Haber

P D Jeffrey

J Novotny

S Sheriff

M N Margolies

Affiliations

Soon will be listed here.

Abstract

Antibody LB4, produced by a spontaneous variant of the murine anti-digoxin monoclonal antibody 26-10, has an affinity for digoxin two orders of magnitude lower than that of the parent antibody due to replacement of serine with phenylalanine at position 52 of the heavy chain variable region (Schildbach, J.F., Panka, D.J., Parks, D.R., et al., 1991, J. Biol. Chem. 266, 4640-4647). To examine the basis for the decreased affinity, a panel of engineered antibodies with substitutions at position 52 was created, and their affinities for digoxin were measured. The antibody affinities decreased concomitantly with increasing size of the substituted side chains, although the shape of the side chains also influenced affinity. The crystal structure of the 26-10 Fab complexed with digoxin (P.D.J., R.K. Strong, L.C. Sieker, C. Chang, R.L. Campbell, G.A. Petsko, E.H., M.N.M., & S.S., submitted for publication) shows that the serine at heavy chain position 52 is not in contact with hapten, but is adjacent to a tyrosine at heavy chain position 33 that is a contact residue. The mutant antibodies were modeled by applying a conformational search procedure to position side chains, using the 26-10 Fab crystal structure as a starting point. The results suggest that each of the substituted side chains may be accommodated within the antibody without substantial structural rearrangement, and that none of these substituted side chains are able to contact hapten. These modeling results are consistent with the substituents at position 52 having only an indirect influence upon antibody affinity.(ABSTRACT TRUNCATED AT 250 WORDS)

Citing Articles

An Outdated Notion of Antibody Specificity is One of the Major Detrimental Assumptions of the Structure-Based Reverse Vaccinology Paradigm, Which Prevented It from Helping to Develop an Effective HIV-1 Vaccine.

van Regenmortel M Front Immunol. 2014; 5:593.

PMID: 25477882 PMC: 4235417. DOI: 10.3389/fimmu.2014.00593.

Basic research in HIV vaccinology is hampered by reductionist thinking.

van Regenmortel M Front Immunol. 2012; 3:194.

PMID: 22787464 PMC: 3391733. DOI: 10.3389/fimmu.2012.00194.

Verdino P, Aldag C, Hilvert D, Wilson I Proc Natl Acad Sci U S A. 2008; 105(33):11725-30.

PMID: 18689687 PMC: 2575270. DOI: 10.1073/pnas.0801783105.

An efficient visualization tool for the analysis of protein mutation matrices.

David M, Lapid C, Daria V BMC Bioinformatics. 2008; 9:218.

PMID: 18442400 PMC: 2390542. DOI: 10.1186/1471-2105-9-218.

26-10 Fab-digoxin complex: affinity and specificity due to surface complementarity.

Jeffrey P, Strong R, Sieker L, Chang C, Campbell R, Petsko G Proc Natl Acad Sci U S A. 1993; 90(21):10310-4.

PMID: 8234291 PMC: 47764. DOI: 10.1073/pnas.90.21.10310.

References

Riechmann L, Clark M, Waldmann H, Winter G . Reshaping human antibodies for therapy. Nature. 1988; 332(6162):323-7. DOI: 10.1038/332323a0. View

Sheriff S, Hendrickson W, Smith J . Structure of myohemerythrin in the azidomet state at 1.7/1.3 A resolution. J Mol Biol. 1987; 197(2):273-96. DOI: 10.1016/0022-2836(87)90124-0. View

Panka D, Parks D, Peterson L, Herzenberg L, Haber E, Margolies M . Variable region framework differences result in decreased or increased affinity of variant anti-digoxin antibodies. Proc Natl Acad Sci U S A. 1988; 85(9):3080-4. PMC: 280147. DOI: 10.1073/pnas.85.9.3080. View

Chien N, Roberts V, Giusti A, Scharff M, Getzoff E . Significant structural and functional change of an antigen-binding site by a distant amino acid substitution: proposal of a structural mechanism. Proc Natl Acad Sci U S A. 1989; 86(14):5532-6. PMC: 297657. DOI: 10.1073/pnas.86.14.5532. View

Padlan E, Silverton E, Sheriff S, Cohen G, Davies D . Structure of an antibody-antigen complex: crystal structure of the HyHEL-10 Fab-lysozyme complex. Proc Natl Acad Sci U S A. 1989; 86(15):5938-42. PMC: 297746. DOI: 10.1073/pnas.86.15.5938. View

Novotny J, Bruccoleri R, Saul F . On the attribution of binding energy in antigen-antibody complexes McPC 603, D1.3, and HyHEL-5. Biochemistry. 1989; 28(11):4735-49. DOI: 10.1021/bi00437a034. View

Herron J, He X, MASON M, Voss Jr E, EDMUNDSON A . Three-dimensional structure of a fluorescein-Fab complex crystallized in 2-methyl-2,4-pentanediol. Proteins. 1989; 5(4):271-80. DOI: 10.1002/prot.340050404. View

Chothia C, Lesk A, Tramontano A, Levitt M, AIR G, Sheriff S . Conformations of immunoglobulin hypervariable regions. Nature. 1989; 342(6252):877-83. DOI: 10.1038/342877a0. View

Padlan E . On the nature of antibody combining sites: unusual structural features that may confer on these sites an enhanced capacity for binding ligands. Proteins. 1990; 7(2):112-24. DOI: 10.1002/prot.340070203. View

10.

Near R, Ng S, Hudson N, Margolies M, Seidman J, Haber E . Heavy and light chain contributions to antigen binding in an anti-digoxin chain recombinant antibody produced by transfection of cloned anti-digoxin antibody genes. Mol Immunol. 1990; 27(9):901-9. DOI: 10.1016/0161-5890(90)90157-u. View

11.

Alzari P, Spinelli S, Mariuzza R, Boulot G, POLJAK R, Jarvis J . Three-dimensional structure determination of an anti-2-phenyloxazolone antibody: the role of somatic mutation and heavy/light chain pairing in the maturation of an immune response. EMBO J. 1990; 9(12):3807-14. PMC: 552147. DOI: 10.1002/j.1460-2075.1990.tb07598.x. View

12.

Near R, Bruccoleri R, Novotny J, Hudson N, White A . The specificity properties that distinguish members of a set of homologous anti-digoxin antibodies are controlled by H chain mutations. J Immunol. 1991; 146(2):627-33. View

13.

Mian I, Bradwell A, Olson A . Structure, function and properties of antibody binding sites. J Mol Biol. 1991; 217(1):133-51. DOI: 10.1016/0022-2836(91)90617-f. View

14.

Schildbach J, Panka D, Parks D, Jager G, Novotny J, Herzenberg L . Altered hapten recognition by two anti-digoxin hybridoma variants due to variable region point mutations. J Biol Chem. 1991; 266(7):4640-7. View

15.

He J, Quiocho F . A nonconservative serine to cysteine mutation in the sulfate-binding protein, a transport receptor. Science. 1991; 251(5000):1479-81. DOI: 10.1126/science.1900953. View

16.

Tempest P, Bremner P, Lambert M, Taylor G, Furze J, Carr F . Reshaping a human monoclonal antibody to inhibit human respiratory syncytial virus infection in vivo. Biotechnology (N Y). 1991; 9(3):266-71. DOI: 10.1038/nbt0391-266. View

17.

Lavoie T, Drohan W . Experimental analysis by site-directed mutagenesis of somatic mutation effects on affinity and fine specificity in antibodies specific for lysozyme. J Immunol. 1992; 148(2):503-13. View

18.

Kettleborough C, Saldanha J, Heath V, Morrison C, Bendig M . Humanization of a mouse monoclonal antibody by CDR-grafting: the importance of framework residues on loop conformation. Protein Eng. 1991; 4(7):773-83. DOI: 10.1093/protein/4.7.773. View

19.

GREENWOOD F, Hunter W, GLOVER J . THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963; 89:114-23. PMC: 1202279. DOI: 10.1042/bj0890114. View

20.

Padlan E, Davies D . Variability of three-dimensional structure in immunoglobulins. Proc Natl Acad Sci U S A. 1975; 72(3):819-23. PMC: 432411. DOI: 10.1073/pnas.72.3.819. View