Cloning, expression and chromosomal localization of human Ca2+/calmodulin-dependent protein kinase kinase |
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| Authors: | Li-Sung Hsu Ann-Ping Tsou Chin-Wen Chi Chen-Hsen Lee Jeou-Yuan Chen |
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| Affiliation: | (1) Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan, ROC;(2) Institute of Biotechnology in Medicine, National Yang-Ming University, Taipei, Taiwan, ROC;(3) Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan, ROC;(4) Department of Medical Research, Veterans General Hospital, Taipei, Taiwan, ROC;(5) Department of Surgery, Veterans General Hospital, Taipei, Taiwan, ROC;(6) Institute of Biomedical Sciences, Academia Sinica, 11529 Taipei, Taiwan (ROC) |
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| Abstract: | A human cDNA clone encoding the calcium/calmodulin-dependent protein kinase kinase (CaMKK) was isolated by RT-PCR amplification of the fragment corresponding to the conserved kinase catalytic domain followed by rapid amplification of cDNA ends and cDNA library screening. Compilation of nucleotide sequencing data yielded a consensus cDNA sequence of 1.9 kb with an open reading frame of 1,251 nucleotides in length which translates to a polypeptide of 417 amino acids (47 kd). It showed significant homology to the rat brain CaMKK isozymes. The human CaMKK, which was expressed as a Flag-tagged protein in human non-small cell lung cancer H-1299 cells followed by immunoprecipitation with anti-Flag antibody, was shown to phosphorylate recombinant human CaMK I in a calcium/CaM-dependent fashion. Northern blot analysis revealed that human CaMKK is ubiquitously expressed, with brain showing the highest level of expression. The CaMKK gene is localized to human chromosome 12. The presence of cDNA clones with divergent 3' terminal sequences suggests a family of CaMKK variants which may arise from alternative splicing. |
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| Keywords: | Ca2+/calmodulin-dependent protein kinase Human CaMKK Cloning Tissue expression Chromosomal location |
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