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Isolation of mitotic p34 cdc2 apoenzyme from human cells
Author(s) -
Meikrantz William,
Feldman Robert P.,
Sladicka Melissa M.,
Ho David,
Krupnick Jason,
Anderson Karen,
Schlegel Robert A.
Publication year - 1991
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(91)81281-c
Subject(s) - biochemistry , mitosis , enzyme , phosphorylation , yeast , cyclin dependent kinase 1 , biology , tyrosine , protein tyrosine phosphatase , microbiology and biotechnology , phosphatase , chemistry , cell cycle , gene
A simple procedure was devised for isolating from homogenates of mitotic cells the human homolog to the fission yeast cdc2 gene product. The identity of the purified protein was established with anti‐p34 cdc2 antibodies and p13 suc1 , both specific ligands for p34 cdc2 . Active‐site labeling with oxidized [α 32 P]ATP showed the purified molecule to be an ATP‐binding protein. Its ability to phosphorylate casein but not histone, and its phosphorylation on tyrosine, detected by anti‐phosphotyrosine antibodies, indicates the form of p34 cdc2 purified is the inactive or apoenzyme form. Purified quantities of human p34 cdc2 should be of considerable value in establishing the mechanism of its activation at mitosis by phosphatases.