Xc. Sun et al., MOLECULAR-CLONING OF CDNA-ENCODING MITOCHONDRIAL VERY-LONG-CHAIN ACYL-COA DEHYDROGENASE FROM BOVINE HEART, Zhongguo yaoli xuebao, 18(1), 1997, pp. 25-32
To clone the cDNA encoding an isoenzyme of mitochondrial very-long-cha
in acyl-CoA dehydrogenase (VLCAD) from bovine heart lambda gt11 and la
mbda gt10 cDNA libraries. METHODS: The clone was isolated with immunos
creening technique and validated by (1) the microsequences of the N-te
rminus and three internal proteolytic fragments from the purified enzy
me; (2) identification of the acyl-CoA dehydrogenase (AD) signature se
quence; and (3) high homology of the deduced peptide sequences, as exp
ected, with those of rat liver mitochondrial VLCAD. RESULTS: The cDNA
(2203 bp) corresponds to a similar to 2.4-kb mRNA band from the same t
issue source revealed by a Northern blotting. The deduced peptide sequ
ence of 655 amino acids (70 537 Da) is composed of a 40-amino acid mit
ochondrial leader peptide moiety (4 346 Da) and a 615-amino acid pepti
de as a mature protein (66 191 Da). A comparison of the peptide sequen
ces in the AD family shows the major diversity in their signal sequenc
es, suggesting a structural basis for their different mitochondrial lo
cations. The catalytic sites are all highly conserved among VLCAD. Ser
-251 analogous to and Cys-215 diversified to other family members. A p
seudo-consensus sequence of leucine zipper was found in the C-terminal
region from Leu-568 to Leu-589, implying a mechanism whereby the dime
r of this protein is formed by zipping these leucine residues from the
alpha-helixes of 2 monomers. CONCLUSION: The isolated cDNA clone enco
des an isoenzyme of mitochondrial VLCAD in bovine heart.