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Cloning, expression and chromosomal localization of human Ca2+/calmodulin-dependent protein kinase kinase

  • Original Paper
  • Published:
Journal of Biomedical Science

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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Authors and Affiliations

  1. Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan, ROC

    Li-Sung Hsu

  2. Institute of Biotechnology in Medicine, National Yang-Ming University, Taipei, Taiwan, ROC

    Ann-Ping Tsou

  3. Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan, ROC

    Chin-Wen Chi

  4. Department of Medical Research, Veterans General Hospital, Taipei, Taiwan, ROC

    Chin-Wen Chi

  5. Department of Surgery, Veterans General Hospital, Taipei, Taiwan, ROC

    Chen-Hsen Lee

  6. Institute of Biomedical Sciences, Academia Sinica, 11529, Taipei, Taiwan (ROC)

    Li-Sung Hsu & Jeou-Yuan Chen

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  1. Li-Sung Hsu
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  2. Ann-Ping Tsou
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  4. Chen-Hsen Lee
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Hsu, LS., Tsou, AP., Chi, CW. et al. Cloning, expression and chromosomal localization of human Ca2+/calmodulin-dependent protein kinase kinase. J Biomed Sci 5, 141–149 (1998). https://doi.org/10.1007/BF02258368

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