Abstract
Monoclonal antibodies (mAbs) were generated that recognize UvrA and UvrB proteins. These proteins are components of the Uvr(A)BC endonuclease, which initiates nucleotide excision repair in Escherichia coli. mAbs, which can be used for probing of structural intermediates of Uvr(A)BC endonuclease functioning, were selected for their ability to: (i) recognize different epitopes; (ii) have a high-affinity for native antigenic protein; (iii) preserve functionality of the Uvr protein in immunocomplex. The adherence of anti-Uvr mAbs with these criteria was verified by additivity and competition tests, and by their influence on the ATPase activities of UvrA and UvrB*, the functionally active proteolytic fragment of UvrB. Two out of twelve anti-UvrA and seven out of thirteen anti-UvrB/anti-UvrB* hybridoma lines were shown to satisfy these criteria. Recognition of UvrA and UvrB deletion mutant proteins by mAbs was used to map their epitopes. Epitopes of A2D1 and A2B1 mAbs were mapped to regions of amino acids 230-281 and 560-680 of UvrA, respectively. Epitopes of anti-UvrB/UvrB* mAbs were assigned to the following amino acid regions of UvrB: B2A1, 8-61; B2C5 and B*2E3, 171-278; B2E2, 631-673; B3C1, 1-7 and/or 62-170; B*2B9, 473-630; B*3E...Continue Reading
References
Oct 1, 1991·Trends in Biochemical Sciences·M E Goldberg
Apr 11, 1986·Nucleic Acids Research·C BackendorfP van de Putte
Nov 11, 1986·Nucleic Acids Research·A T YeungL Grossman
May 6, 1986·Biochemistry·L Djavadi-OhanianceM E Goldberg
Mar 18, 1985·Journal of Immunological Methods·B FriguetM E Goldberg
Jun 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·T P Hopp, K R Woods
Jun 10, 1983·Journal of Immunological Methods·B FriguetM Goldberg
Jul 1, 1984·Molecular Immunology·B FriguetM E Goldberg
Mar 3, 1995·The Journal of Biological Chemistry·N E ThompsonR R Burgess
Jul 7, 1995·The Journal of Biological Chemistry·A Sancar
Feb 3, 1993·Journal of Immunological Methods·B FriguetL Djavadi-Ohaniance
Apr 7, 1995·The Journal of Biological Chemistry·D S HsuA Sancar
Jul 22, 1994·Journal of Molecular Biology·G F MoolenaarP van de Putte
Oct 29, 1993·Gene·L LiS Chandrasegaran
Dec 22, 1995·The Journal of Biological Chemistry·G F MoolenaarN Goosen
May 10, 1996·The Journal of Biological Chemistry·A W Karzai, R McMacken
Aug 2, 1996·The Journal of Biological Chemistry·J MarszalekJ M Kaguni