Comparison of two cytochromes P-450 from Candida maltosa: primary structures, substrate specificities and effects of their expression in Saccharomyces cerevisiae on the proliferation of the endoplasmic reticulum.
cDNAs were cloned, sequenced and expressed which encode two different cytochrome P-450 forms of the alkane-assimilating yeast Candida maltosa, designated as P-450Cm1 and P-450Cm2. The amino acid sequences deduced were about 55% identical. Expression in Saccharomyces cerevisiae resulted in the formation of intact microsomal P-450 systems catalyzing the hydroxylation of n-hexadecane and lauric acid with significantly different substrate preferences. A massive proliferation of the endoplasmic reticulum was observed in the S. cerevisiae cells which produced P-450. Depending on the P-450 form expressed, distinctly organized stacks of paired membranes appeared and occupied considerable areas of the cytoplasm. As shown by immunoelectron microscopy for P-450Cm1, the protein expressed was highly concentrated within these newly formed membrane structures.
Central Institute of Molecular Biology, Berlin-Buch/Federal Republic of Germany.
W H
WH
Schunck
F
F
Vogel
B
B
Gross
E
E
Kärgel
S
S
Mauersberger
K
K
Köpke
C
C
Gengnagel
H G
HG
Müller
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D002175:Candida / Q000201:enzymology*; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000235:genetics; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D015870:Gene Expression; D005800:Genes, Fungal; D006900:Hydroxylation; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D008854:Microscopy, Electron; D008861:Microsomes / Q000201:enzymology; D008969:Molecular Sequence Data; D005810:Multigene Family; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000648:ultrastructure; D016415:Sequence Alignment; D013379:Substrate Specificity
D007527:Isoenzymes; D004247:DNA; D003577:Cytochrome P-450 Enzyme System