Format

Send to

Choose Destination
Proc Natl Acad Sci U S A. 2005 Jun 7;102(23):8114-9. Epub 2005 May 31.

Building a human kinase gene repository: bioinformatics, molecular cloning, and functional validation.

Author information

1
Harvard Institute of Proteomics, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 320 Charles Street, Cambridge, MA 02141, USA.

Abstract

Kinases catalyze the phosphorylation of proteins, lipids, sugars, nucleosides, and other important cellular metabolites and play key regulatory roles in all aspects of eukaryotic cell physiology. Here, we describe the mining of public databases to collect the sequence information of all identified human kinase genes and the cloning of the corresponding ORFs. We identified 663 genes, 511 encoding protein kinases, and 152 encoding nonprotein kinases. We describe the successful cloning and sequence verification of 270 of these genes. Subcloning of this gene set in mammalian expression vectors and their use in high-throughput cell-based screens allowed the validation of the clones at the level of expression and the identification of previously uncharacterized modulators of the survivin promoter. Moreover, expressions of the kinase genes in bacteria, followed by autophosphorylation assays, identified 21 protein kinases that showed autocatalytic activity. The work described here will facilitate the functional assaying of this important gene family in phenotypic screens and their use in biochemical and structural studies.

PMID:
15928075
PMCID:
PMC1149441
DOI:
10.1073/pnas.0503141102
[Indexed for MEDLINE]
Free PMC Article

Publication type, MeSH terms, Substances, Secondary source ID

Publication type

MeSH terms

Substances

Secondary source ID

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center