Mo. James et al., CDNA AND PROTEIN-SEQUENCE OF A MAJOR FORM OF P450, CYP2L, IN THE HEPATOPANCREAS OF THE SPINY LOBSTER, PANULIRUS-ARGUS, Archives of biochemistry and biophysics, 329(1), 1996, pp. 31-38
A P450 fraction was previously isolated from spiny lobster hepatopancr
eas microsomes and shown in reconstitution experiments to be efficient
in catalyzing the monooxygenation of benzphetamine, aminopyrine, benz
o(a)pyrene, progesterone, and testosterone. In this study, N-terminal
sequence information up to residue 39 was obtained from this P450 and
used to design degenerate primers for screening a cDNA library constru
cted from hepatopancreas mRNA. Clones were obtained that contained par
t of the coding region of a P450 protein. Further exact primers were d
esigned that permitted the isolation of clones containing coding infor
mation for other parts of the P450 sequence, as well as a clone that c
oded for the complete P450 protein sequence, The open reading frame of
the complete coding region corresponded to a protein of 492 amino aci
ds. The deduced amino acid sequence of this P450 was about 36% similar
to individual mammalian P450s in the 2 family and did not show strong
matches with other proteins in the data base, Based on sequence and t
he previously determined function, this spiny lobster P450 was assigne
d by the P450 nomenclature committee to a new P450 subfamily, CYP2L. T
his is the first description of a P450 primary sequence from a marine
crustacean species and the first assignment of an invertebrate P450 in
to the 2 family. (C) 1996 Academic Press, Inc.