Expression, purification and kinetic behaviour of fission yeast low Mrprotein‐tyrosine phosphatase

FEBS Letters

Raugei, Giovanni¹  Magherini, Francesca¹  Manao, Giampaolo¹  Modesti, Alessandra¹  Ramponi, Giampietro¹  Camici, Guido¹  Cirri, Paolo¹  Carraresi, Laura¹ 

[1] Dipartimento di Scienze Biochimiche, Università di Firenze, Viale Morgagni 50, 50134 Firenze, Italy

关键词: Phosphotyrosine protein phosphatase;    Fission yeast;    Active site;    stp1 +;    small tyrosine phosphatase);    LMW-PTP;    low molecular weight phosphotyrosine protein phosphatase;    PTPases;    phosphotyrosine protein phosphatases;    PTK;    phosphotyrosine protein kinase;    kDa;    kiloDalton;    cGMP;    cyclic guanosine monophosphate;    IF1 (isoform 1);    IF2 (isoform 2);    GST (glutathione S-transferase);    IPTG;    isopropylthiogalactoside;    pNPP;    p-nitrophenylphosphate;    l-P-Tyr;    l-phosphotyrosine;    SDS-PAGE;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;    PCR;    polymerase chain reaction;   

DOI  :  10.1016/0014-5793(95)01220-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A gene named stp1 ⁺, coding for a 17.5-kDa protein, that rescues cdc25-22 when overexpressed, has been previously isolated from fission yeast. Here we describe the expression and purification of Stp1 protein as a fusion with the glutathione S-transferase in E. coli and its kinetic characterisation. Stp1 deduced protein sequence shows an high homology to members of a class of cytosolic low M _r protein phosphatase previously known to exist only in mammalian species. Stp1 has a kinetic behaviour that appears to be intermediate with respect to the two isoenzymatic forms of low M _r protein tyrosine phosphatases present in mammalian tissues. These differing kinetic characteristics are mainly due to the sequence 45–56 that is spatially close to the active site pocket.

【 授权许可】

Unknown