3-Hydroxy-3-methylglutaryl-coenzyme A reductase of the sea urchin embryo. Deduced structure and regulatory properties.
3-Hydroxy-3-methylglutaryl-coenzyme A reductase activity is developmentally regulated in the sea urchin Strongylocentrotus purpuratus (Woodward, H. D., Allen, J. M. C., and Lennarz, W. J. (1988) J. Biol. Chem. 263, 2513-2517). To study the structural and regulatory properties of this enzyme, we isolated and sequenced a 3-kb cDNA encoding the sea urchin embryo reductase. The deduced amino acid sequence of this cDNA predicted a protein structure consisting of a hydrophobic N-terminal region containing seven potential membrane-spanning domains and a somewhat less hydrophobic C-terminal domain joined by a hydrophilic linker region. Comparison with reductase from mammalian sources revealed that the N-terminal membrane domain and the C-terminal cytoplasmic domain exhibited high sequence similarity, whereas the domain that linked these two showed little or no sequence similarity. We investigated the possibility that sterols or sterol derivatives might be involved in the marked change that occurs in the level of reductase activity over development. Enzyme activity and reductase mRNA levels measured in extracts from embryos cultured in the presence of cholesterol, 25-hydroxycholesterol, dolichol, or mevalonic acid were found to be virtually unchanged as compared to control embryos. Similar experiments with mevinolin, a competitive inhibitor of reductase, failed to show a drug-induced change in enzyme or mRNA level. Thus, despite structural similarities the sea urchin embryo enzyme differs markedly from the mammalian enzyme with respect to regulation, since its level is neither repressed by sterols nor induced by mevinolin. Moreover, it appears unlikely that sterols or sterol derivatives play a role in the striking change in the level of this enzyme that occurs during development.
Woodward
H D
HD
Department of Biochemistry and Molecular Biology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Allen
J M
JM
Lennarz
W J
WJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004625:Embryo, Nonmammalian / Q000201:enzymology*; D006903:Hydroxymethylglutaryl CoA Reductases / Q000235:genetics / Q000378:metabolism*; D008861:Microsomes / Q000201:enzymology; D008969:Molecular Sequence Data; D011487:Protein Conformation; D015183:Restriction Mapping; D012617:Sea Urchins / Q000196:embryology*
D004247:DNA; D006903:Hydroxymethylglutaryl CoA Reductases