J. Biochem, 1998, Vol. 123, No. 1 47-54
© 1998 Japanese Biochemical Society
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Cloning and Expression of Human Adenylate Kinase 2 Isozymes: Differential Expression of Adenylate Kinase1 and 2 in Human Muscle Tissues1
*Molecular and Cellular Biology Research Division, Korea Research Institute of Bioscience and Biotechnology Taejon 305-333, Korea
Department of Biochemistry, Hanyang University Ansan 425-791, Korea
Department of Surgery, Hanyang University Medical School Seoul 133-792, Korea
§Department of Biological Sciences, KAIST Taijon 305-701, Korea
2To whom correspondence should be addressed at: P.O. Box 115, Yusung, Taejon, 305-600, Korea. Tel: +82-42-860-4180, Fax: + 8242-860-4593, E-mail: choemcbg{at}kribb4680.kribb.re.kr
A cDNA clone coding for adenylate kinase 2B was isolated from fetal liver, and the expression of AK2 was investigated in human tissues. The ORF in the cDNA clone for human AK2B predicted a protein comprising 232 amino acids (25.6 kDa). The features of AK2A and AK2B sequences in human were the same as those in the bovine system. Each of the recombinant proteins, AK2A and AK2B, was expressed in Escherichia coli cells, and the purified recombinant proteins were enzymatically active. The distribution of AK2 transcripts in various human tissues was examined by Northern analysis. Unlike in the bovine system, it was found that the AK2A transcript was the major form of AK2 mRNA species in all human tissues. The transcripts of AK2 isozymes were relatively abundant in heart, liver, and also in skeletal muscle, where the expression level of AK2 was known to be low. Western blot analysis of AK isozymes in human heart and skeletal muscle revealed that AK2 protein was found only in heart, whereas AK1 was detected in both tissues. These tissue-specific expressions of the AK isozymes in human might suggest the presence of organ-specific regulation of the AK2 gene including a post-transcriptional control in skeletal muscle.
1This work was supported by a grant from the Ministry of Science and Technology (HAN Project HS050M). The nucleotide sequence data reported in this paper will appear in the EMBL, DDBJ, and GenBank nucleotide sequence databases with the accession number U39945 [GenBank] (human adenylate kinase 2A) and U54645 [GenBank] (human adenylate kinase 2B).
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