Nucleotide sequence for cDNA of bovine mitochondrial ATP-dependent protease and determination of N-terminus of the mature enzyme from the adrenal cortex.
We have determined the cDNA sequence encoding bovine mitochondrial ATP-dependent Lon protease. Since the 5'-end region of the cDNA was highly GC-rich and thus could not be amplified by the 5'-RACE method, a genomic DNA fragment containing an in-frame ATG was isolated and sequenced. The translated amino acid sequence contained 961 amino acids with a calculated molecular weight 106,665. Sequence similarities of the bovine enzyme to human and E. coli orthologs were 92 and 27%, respectively. The N-terminal amino acid sequence seemed to be a mitochondrial targeting signal. To determine the cleavage site of the signal sequence we analyzed the mature enzyme purified from bovine adrenocortical mitochondria. Analysis of CNBr-digested peptides revealed that the N-terminus was heterogeneous. We suggest that nonspecific aminopeptidase might remove several amino acids from the N-terminus after mitochondrial processing peptidase has cleaved Gly(67)-Leu(68) or Leu(68)-Trp(69).
Yamamoto
Misa
M
Laboratory Sciences, Faculty of Health Sciences,Yamaguchi University School of Medicine, Ube 755-8505, Japan.
Hiroi
Tomoko
T
Kohno
Hiroyuki
H
Yamamoto
Yoshimi
Y
Hara
Masayuki
M
Tanaka
Tatehiko
T
Mamba
Kouichi
K
Watabe
Shoji
S
D049069:ATP-Dependent Proteases / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D000302:Adrenal Cortex / Q000201:enzymology*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D013058:Mass Spectrometry; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D010446:Peptide Fragments; D049069:ATP-Dependent Proteases