The previously isolated cDNA encoding human adenylate kinase (AK) isozyme 3 was recently renamed AK4. Consequently, human AK3 cDNA remains to be identified and we have little information about the functional relationship between human AK3 and AK4. In pursuit of the physiological roles of both the AK3 and AK4 proteins, we first isolated an authentic human AK3 cDNA and compared their expression. Nucleotide sequencing revealed that the cDNA encoded a 227-amino-acid protein, with a deduced molecular mass of 25.6kDa, that shares greater homology with the AK3 cDNAs isolated from bovine and rat than that from human. We named the isolated cDNA AK3. Northern-blot analysis revealed that AK3 mRNA was present in all tissues examined, and was highly expressed in heart, skeletal muscle and liver, moderately expressed in pancreas and kidney, and weakly expressed in placenta, brain and lung. On the other hand, we found that human AK4 mRNA was highly expressed in kidney, moderately expressed in heart and liver and weakly expressed in brain. Western-blot analysis demonstrated expression profiles of AK3 and AK4 that were similar to their mRNA expression patterns in each tissue. Over expression of AK3, but not AK4, in both Escherichia coli CV2, a temperature-sensitive AK mutant, and a human embryonic kidney-derived cell line, HEK-293, not only produced significant GTP:AMP phosphotransferase (AK3) activity, but also complemented the CV2 cells at 42°C. Subcellular and submitochondrial fractionation analysis demonstrated that both AK3 and AK4 are localized in the mitochondrial matrix.
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August 2001
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Research Article|
August 08 2001
Structure and expression of human mitochondrial adenylate kinase targeted to the mitochondrial matrix
Takafumi NOMA;
Takafumi NOMA
1
∗Department of Biochemistry, Yamaguchi University School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan
1To whom correspondence should be addressed (e-mail tnoma@po.cc.yamaguchi-u.ac.jp).
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Koichi FUJISAWA;
Koichi FUJISAWA
∗Department of Biochemistry, Yamaguchi University School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan
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Yasuhiro YAMASHIRO;
Yasuhiro YAMASHIRO
∗Department of Biochemistry, Yamaguchi University School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan
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Miho SHINOHARA;
Miho SHINOHARA
∗Department of Biochemistry, Yamaguchi University School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan
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Atsushi NAKAZAWA;
Atsushi NAKAZAWA
∗Department of Biochemistry, Yamaguchi University School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan
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Toshikazu GONDO;
Toshikazu GONDO
†Department of Pathology, Yamaguchi University School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan
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Tokuhiro ISHIHARA;
Tokuhiro ISHIHARA
†Department of Pathology, Yamaguchi University School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505, Japan
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Kumiko YOSHINOBU
Kumiko YOSHINOBU
‡Gene Technology Center, Kumamoto University, 2-2-1, Honjo, Kumamoto, 860-0811, Japan
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Publisher: Portland Press Ltd
Received:
May 15 2001
Accepted:
June 12 2001
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2001
2001
Biochem J (2001) 358 (1): 225–232.
Article history
Received:
May 15 2001
Accepted:
June 12 2001
Citation
Takafumi NOMA, Koichi FUJISAWA, Yasuhiro YAMASHIRO, Miho SHINOHARA, Atsushi NAKAZAWA, Toshikazu GONDO, Tokuhiro ISHIHARA, Kumiko YOSHINOBU; Structure and expression of human mitochondrial adenylate kinase targeted to the mitochondrial matrix. Biochem J 15 August 2001; 358 (1): 225–232. doi: https://doi.org/10.1042/bj3580225
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