Isolation and characterization of microplasminogen. A low molecular weight form of plasminogen.
A functionally active human microplasminogen without kringle structures was produced by incubation of plasminogen with urokinase-free plasmin at an alkaline pH. The microplasminogen was purified by affinity chromatography on lysine- and soybean trypsin inhibitor-Sepharose and by chromofocusing. Human plasminogen is specifically cleaved at Arg529-Lys530 by plasmin to form microplasminogen, which consists of a single polypeptide of 261 residues from the COOH-terminal portion of native plasminogen. It has an Mr of 28,617, calculated from the sequence, which is consistent with the molecular weight determined by sodium dodecyl sulfate gel electrophoresis. Microplasminogen is a slightly basic protein and is eluted from a chromofocusing column at pH 8.3. It can be activated by urokinase and streptokinase to a catalytically active microplasmin. The specific amidolytic activity of microplasmin is about three times higher than Lys77-plasmin on a weight basis and is about the same on a molar basis. The activation of microplasminogen by streptokinase is slower than that of either Glu-plasminogen or Lys77-plasminogen. On the other hand, the activation of microplasminogen by urokinase is faster than that of either of the latter. The Arg560-Val561 bond is cleaved during activation of both microplasminogen and native plasminogen.
Shi
G Y
GY
Biochemistry Department, Medical College, National Cheng-Kung University, Tainan, Taiwan, Republic of China.
Wu
H L
HL
D000595:Amino Acid Sequence; D001120:Arginine; D002851:Chromatography, High Pressure Liquid; D005341:Fibrinolysin / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008239:Lysine; D008832:Microchemistry; D008969:Molecular Sequence Data; D015394:Molecular Structure; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis / Q000302:isolation & purification*; D010958:Plasminogen / Q000032:analysis / Q000302:isolation & purification*
D010446:Peptide Fragments; D010958:Plasminogen; D001120:Arginine; D005341:Fibrinolysin; D008239:Lysine