The construction and characterization of human pro-urokinase mutant / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 573-578, 2005.
Artigo
em Chinês
| WPRIM
| ID: wpr-305200
ABSTRACT
It is very easy for the pro-UK to lose it's biological activity because of the digestion of pro-UK by the thrombin or the inhibition of pro-UK by the PAI-I. So three pro-UK mutant (pro-UK) genes were constructed in this experiment with the PCR point-mutant method. The thrombin cleavage site Arg156 in pro-UK was mutated into His156, and named as pro-UKM1; PAI binding sites Arg178, Arg179, Arg181 were mutated into Lys178, Lys179, His181, named as pro-UKM2; The mutant containing His156, Lys178, Lys179, His181 as pro-UKM3. Three mutants were expressed in CHO cells respectively and analyzed with SDS-PAGE fibrin plate assay and western blot. The results showed that the three mutants and the native pro-UK have the same single electrophoresis band indicating most of the pro-UK was single chain. In vitro plasma clot lysis assays indicated that the pro-UKM1 have the ability to resistant against thrombin digestion; pro-UK2 could resist against PAI inhibition; while pro-UK3 improved resistances against both thrombin and PAI. It looks very promising that the pro-UK3 can be a new medicine of dissolving thrombus.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Proteínas Recombinantes
/
Dados de Sequência Molecular
/
Sequência de Bases
/
Transfecção
/
Ativador de Plasminogênio Tipo Uroquinase
/
Western Blotting
/
Cricetulus
/
Células CHO
/
Clonagem Molecular
/
Eletroforese em Gel de Poliacrilamida
Limite:
Animais
/
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2005
Tipo de documento:
Artigo
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