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Pull-down of Calmodulin-binding Proteins
07:51

Pull-down of Calmodulin-binding Proteins

Published on: January 23, 2012

cDNA cloning and expression of human calmodulin-dependent protein kinase IV

T Kitani1, S Okuno, H Fujisawa

  • 1Department of Biochemistry, Asahikawa Medical College, Hokkaido.

Journal of Biochemistry
|April 1, 1994
PubMed

Insights

Researchers isolated and sequenced a human cDNA clone for Calmodulin-dependent protein kinase IV (CaM-kinase IV). This protein kinase, abundant in the brain, showed high sequence identity to its rat counterpart.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Neuroscience

Background:

  • Calmodulin-dependent protein kinase IV (CaM-kinase IV) is a calcium-responsive enzyme.
  • It is notably abundant in the brain and thymus.

Purpose of the Study:

  • To isolate and characterize the human cDNA encoding CaM-kinase IV.
  • To determine the nucleotide and amino acid sequences of human CaM-kinase IV.
  • To compare the human sequence with its rat ortholog.

Main Methods:

  • Isolation of a human cDNA clone from a Jurkat cell library.
  • Nucleotide sequencing of the cDNA.
  • Western blot analysis using a polyclonal antibody against recombinant human CaM-kinase IV.

Main Results:

  • A human cDNA clone encoding a 473-amino acid protein (51,925 Da) was identified.
  • The human sequence exhibited 81% nucleotide and 80% amino acid identity to the rat enzyme.
  • Western blots detected two human CaM-kinase IV isoforms (60 and 61 kDa) in Jurkat cells and cross-reacted with rat isoforms (62 and 64 kDa).

Conclusions:

  • The study successfully cloned and sequenced human CaM-kinase IV.
  • Significant sequence homology exists between human and rat CaM-kinase IV.
  • The findings provide molecular insights into CaM-kinase IV structure and potential isoforms.

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