The Protein Journal

, Volume 27, Issue 1, pp 43–49 | Cite as

Cytoplasmic and Mitochondrial Creatine Kinases from the Skeletal Muscle of Sperm Whale (Physeter macrocephalus). Molecular Cloning and Enzyme Characterization

  • Kentaro Iwanami
  • Kouji Uda
  • Hiroshi Tada
  • Tomohiko Suzuki


We have amplified two cDNAs, coding for creatine kinases (CKs), from the skeletal muscle of sperm whale Physeter macrocephalus by PCR, and cloned these cDNAs into pMAL plasmid. These are the first CK cDNA and deduced amino acid sequences from cetaceans to be reported. One of the two amino acid sequences is a cytoplasmic, muscle-type isoform (MCK), while the other was identified as a sarcomeric, mitochondrial isoform (sMiCK) that included a mitochondrial targeting peptide. The amino acid sequences of sperm whale MCK and sMiCK showed 94–96% sequence identity with corresponding isoforms of mammalian CKs, and all of the key residues necessary for CK function were conserved. The phylogenetic analyses of vertebrate CKs with three independent methods (neighbor-joining, maximum-likelihood and Bayes) supported the clustering of sperm whale MCK with Bos and Sus MCKs, in agreement with the contemporary view that these groups are closely related. Sperm whale MCK and sMiCK were expressed in Escherichia coli as a fusion protein with maltose-binding protein, and the kinetic constants (K m, K d and k cat) were determined for the forward reaction. Comparison of kinetic constants with those of human and mouse CKs indicated that sperm whale MCK has a comparable affinity for creatine (K m Cr  = 9.38 mM) to that of human MCK, and the sMiCK has two times higher affinity for creatine than the human enzyme. Both the MCK and sMiCK of sperm whale display a synergistic substrate binding (K d /K m = 3.1–7.8) like those of other mammalian CKs.


Creatine kinase Mitochondrial Phosphagen kinase Guanidino kinase Kinetic constant Cetacean Sperm whale Physeter macrocephalus 



arginine kinase


creatine kinase


muscle-type isoform of CK


brain-type isoform of CK


sarcomeric mitochondrial isoform of CK


ubiquitous mitochondrial isoform of CK


creatine; MBP, maltose-binding protein



This work was supported by a Grant-in-Aid for Scientific Research in Japan to TS (17570062).


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Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Kentaro Iwanami
    • 1
  • Kouji Uda
    • 1
  • Hiroshi Tada
    • 1
  • Tomohiko Suzuki
    • 1
  1. 1.Laboratory of Biochemistry, Faculty of ScienceKochi UniversityKochiJapan

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