Factor H is a dermatan sulfate-binding protein: identification of a dermatan sulfate-mediated protease that cleaves factor H.
Dermatan sulfate mediates the blood coagulation cascade by binding to heparin cofactor II and potentiating the antithrombin activity. In order to explore another function of dermatan sulfate, a dermatan sulfate affinity column was prepared from biotinylated dermatan sulfate and Streptavidin Sepharose. When human plasma was applied on the dermatan sulfate column, factor H was bound and cleaved. The cleavage products, a 30-kDa N-terminal fragment and a 120-kDa fragment, were eluted from the column with 500 mM NaCl and detected after Western blotting with anti-factor H. The bond between the tandem arginine residues in the sixth domain of factor H was cleaved. When purified factor H was applied on the column, the factor H was not cleaved and was recovered from the column as an intact 150-kDa fraction. The finding that dermatan sulfate-mediated cleavage of factor H was inhibited by (p-amidinophenyl) methanesulfonyl fluoride, but not N-ethylmaleimide or EDTA, indicates that a serine protease in the plasma was activated on the dermatan sulfate column and factor H was cleaved without intervention of the plasma protease inhibitors. Amidase activity was detected in the effluent from the dermatan sulfate column but was abolished by pretreatment of the plasma with dermatan sulfate. Therefore, dermatan sulfate participates in the activation of a protease as well as having the protease inhibitory action.
Saito
Akio
A
Department of Biochemistry, School of Medicine, Kinki University, 377-2 Ohno-Higashi, Osaka-Sayama, Osaka, 589-8511. asaito@med.kindai.ac.jp
Munakata
Hiroshi
H
D000581:Amidohydrolases / Q000097:blood* / Q000378:metabolism; D017242:Complement Factor H / Q000378:metabolism*; D003871:Dermatan Sulfate / Q000097:blood / Q000378:metabolism*; D006801:Humans; D010664:Phenylmethylsulfonyl Fluoride / Q000031:analogs & derivatives* / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000097:blood* / Q000187:drug effects / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology
C493495:CFH protein, human; D015842:Serine Proteinase Inhibitors; D003871:Dermatan Sulfate; D010664:Phenylmethylsulfonyl Fluoride; C027226:(4-amidinophenyl)methanesulfonyl fluoride; D017242:Complement Factor H; D012697:Serine Endopeptidases; D000581:Amidohydrolases