Komatsu Y, Misawa S, Sukesada A, Ohba Y, Hayashi H
Pharmaceuticals & Biotechnology Laboratory, Nikko Kyodo Co., Ltd., Saitama, Japan.
Biochem Biophys Res Commun. 1993 Oct 29;196(2):773-9. doi: 10.1006/bbrc.1993.2316.
To elucidate the differential roles of N- and C-terminal halves of hirudins in thrombin inhibition, we produced novel recombinant hirudin analogs, CX-397 and CX-397R, having a hybrid amino acid sequence of hirudin variants-1 (HV-1) and -3 (HV-3). CX-397 is composed of the N-terminal half of HV-1 (HV-11-36) combined with the C-terminal half of HV-3 (HV-337-66). CX-397R is the opposite combination. Their anti-thrombin activity was determined by a fluorogenic enzyme assay and compared with that of recombinant HV-1 (rHV-1) and rHV-3. The order of the magnitude of dissociation constants (Ki) of these four hirudin analogs in thrombin inhibition was as follows: CX-397R (0.294 pM) > rHV-1 (0.148 pM) > rHV-3 (0.0593 pM) > CX-397 (0.0433 pM), indicating that CX-397 is the strongest inhibitor among them.
为阐明水蛭素N端和C端在抑制凝血酶中的不同作用，我们制备了新型重组水蛭素类似物CX - 397和CX - 397R，它们具有水蛭素变体-1（HV - 1）和-3（HV - 3）的杂合氨基酸序列。CX - 397由HV - 1的N端部分（HV - 11 - 36）与HV - 3的C端部分（HV - 337 - 66）组合而成。CX - 397R则是相反的组合。通过荧光酶法测定它们的抗凝血酶活性，并与重组HV - 1（rHV - 1）和rHV - 3进行比较。这四种水蛭素类似物在抑制凝血酶时解离常数（Ki）大小顺序如下：CX - 397R（0.294 pM）> rHV - 1（0.148 pM）> rHV - 3（0.0593 pM）> CX - 397（0.0433 pM），表明CX - 397是其中最强的抑制剂。
