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Purification and crystallization of the CDK‐associated protein phosphatase KAP expressed in escherichia coli
Author(s) -
Hanlon Neil,
Barford David
Publication year - 1998
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560070233
Subject(s) - cyclin dependent kinase , escherichia coli , phosphatase , recombinant dna , microbiology and biotechnology , cyclin , biology , kinase , chemistry , biochemistry , enzyme , cell cycle , cell , gene
The kinase associated phosphatase (KAP) is a human dual specificity protein phosphatase that dephosphorylates the cell cycle control protein, cyclin dependent kinase‐2 on Thr 160 in a cyclin dependent manner (Poon & Hunter, 1995). We report here the over‐expression of KAP in Escherichia coli as an N‐terminal His‐tagged protein using a modified pET‐28a T7‐expression vector. The recombinant protein was purified to homogeneity and crystallized. The crystals diffract to 2.3 Ǎ resolution when exposed to synchrotron radiation and belong to space group P6 1 22, or its enantiomorph P6 5 22, with unit cell dimensions a = b 74.5 Ǎ, c = 139.5 Ǎ.