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Buchner, Johannes

Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board Member: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Sies, Helmut / Turk, Boris / Wittinghofer, Alfred


SCImago Journal Rank (SJR) 2015: 1.607
Source Normalized Impact per Paper (SNIP) 2015: 0.751
Impact per Publication (IPP) 2015: 2.609

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1437-4315
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Mutational Analysis of Two Stefin A Epitopes

N. Kopitar-Jerala / R. Jerala / B. Turk / F. Gubensek / V. Turk

Citation Information: Biological Chemistry. Volume 380, Issue 6, Pages 723–726, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/BC.1999.091, June 2005

Publication History

Published Online:
2005-06-01

Abstract

Stefin A, an intracellular inhibitor of cysteine proteinases, is expressed most abundantly in epithelial cells and in cells of lymphatic origin. In order to study its role in normal and pathological conditions we have prepared and characterized monoclonal antibodies against recombinant stefin A. Two high affinity monoclonal antibodies (mAbs) (A22 and C52) were tested for binding to free and papain-complexed stefin A and to a chimeric inhibitor, consisting of 61 amino acid residues of stefin A and 37 carboxy-terminal residues of stefin B. mAb A22 recognized not only free stefin A but also stefin A in complex with papain. The mAbs were further tested for their cross-reactivity against stefin A and B isolated from different mammalian species. On the basis of sequence similarity and tertiary structure of human stefin A we have prepared three mutants–Glu33Lys, Asp61Gly and Asn62Tyr–and their reactivity with the mAbs was tested. The binding affinities of mAb A22 for the Asp61Gly and Asn62Tyr mutants were significantly lower, indicating that the two amino acids are part of the stefin A epitope recognized by A22. The binding of both mAbs to the mutants Gly4Arg and Gly4Glu was comparable to wild-type stefin A.

Citing Articles

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[1]
X. Fan, N. Kopitar-Jerala, A. Premzl, M. Bestagno, O. Burrone, and J. Kos
Biological Chemistry, 2002, Volume 383, Number 11
[2]
Natasa Kopitar-Jerala, Franc Gubenek, and Vito Turk
Biological Chemistry, 2000, Volume 381, Number 12

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