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Cloning of the complete coding region for human protein phosphatase inhibitor 2 using the two hybrid system and expression of inhibitor 2 in E. coli
Author(s) -
Helps Nicholas R.,
Street Alasdair J.,
Elledge Stephen J.,
Cohen Patricia T.W.
Publication year - 1994
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(94)80179-7
Subject(s) - cloning (programming) , chemistry , phosphatase , coding region , escherichia coli , microbiology and biotechnology , biochemistry , computational biology , biology , gene , computer science , enzyme , programming language
The yeast two hybrid system has been employed to identify cDNAs encoding proteins which interact with the γ 1 , isoform of human protein phosphatase 1. Here we report the isolation of a cDNA encoding human protein phosphatase inhibitor 2. The deduced human sequence of 205 amino acids shows 92% identity to inhibitor 2 from rabbit. Human inhibitor 2 was expressed in E. coli and purified to homogeneity. The expressed human protein inhibited both native and bacterially expressed PP1, with the same K i (1 nM) as inhibitor 2 purified from skeletal muscle. A gene or pseudogene for inhibitor 2 may be present near the major histocompatibility complex on chromosome 6.