Premium
Expression, purification and kinetic behaviour of fission yeast low M r protein‐tyrosine phosphatase
Author(s) -
Modesti Alessandra,
Cirri Paolo,
Raugei Giovanni,
Carraresi Laura,
Magherini Francesca,
Manao Giampaolo,
Camici Guido,
Ramponi Giampietro
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01220-9
Subject(s) - protein tyrosine phosphatase , biology , phosphatase , yeast , tyrosine , cytosol , biochemistry , cdc25 , fusion protein , microbiology and biotechnology , peptide sequence , enzyme , gene , recombinant dna , cyclin dependent kinase 1 , cell cycle
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.