Calcium modulates plasmin cleavage of the fibrinogen D fragment gamma chain N-terminus: mapping of monoclonal antibody J88B to a plasmin sensitive domain of the gamma chain.
Plasmin sensitive sites are found on the A alpha, B beta and gamma chains of fibrinogen at regions joining the two C-terminal D fragments with the central E fragment. We have developed a monoclonal antibody (MoAb) reactive with this plasmin sensitive region on the human fibrinogen gamma chain and mapped its epitope. MoAb J88B reacts with gamma chains of both native as well as with reduced and denatured fibrinogen and fibrin, the CNBr fragment of the fibrinogen central domain, plasmin cleaved fragments D, gamma-gamma dimers, but not with plasmic fragments E. These data indicate that J88B maps to the plasmin sensitive domain localized to gamma 63-78. MoAb J88B failed to react with synthetic peptide gamma 70-78, which suggests that the epitope includes the newly exposed N-terminal residues gamma 63-70 of the early plasmic fragment D1A. As calcium has a marked influence on plasmin cleavage of C-terminal sites on the gamma chain, the effects of calcium on modulating plasmin cleavage of D1A to D1 were assessed in the absence or presence of J88B. The results indicated that calcium delays and J88B (+/- calcium) protects the gamma chain from plasmin cleavage at the N-terminus of D1A, suggesting that this enzymatically labile site is calcium-sensitive. Thus, MoAb J88B should prove useful in studies examining the structure of plasmin cleaved fibrinogen and fibrin.
Odrljin
T M
TM
Department of Medicine, University of Rochester School of Medicine and Dentistry, NY 14642, USA.
Rybarczyk
B J
BJ
Francis
C W
CW
Lawrence
S O
SO
Hamaguchi
M
M
Simpson-Haidaris
P J
PJ
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D001665:Binding Sites; D015153:Blotting, Western; D002118:Calcium / Q000494:pharmacology*; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry; D005337:Fibrin / Q000378:metabolism; D005338:Fibrin Fibrinogen Degradation Products / Q000737:chemistry / Q000378:metabolism*; D005341:Fibrinolysin / Q000378:metabolism*; D005343:Fibrinolytic Agents / Q000378:metabolism; D006801:Humans; D011485:Protein Binding
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005338:Fibrin Fibrinogen Degradation Products; D005343:Fibrinolytic Agents; C015214:fibrinogen D fragment; D005337:Fibrin; D004492:Edetic Acid; D005341:Fibrinolysin; D002118:Calcium