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Cloning and sequencing of chloroperoxidase cDNA.
An oligod-d(T) 12-18 primed cDNA library has been prepared from Caldariomyces fumago mRNA. A clone containing a full-length insert was sequenced on the supercoiled plasmid, pBR322. The complete primary sequence of chloroperoxidase has been derived. We have also determined about 73% of the peptide sequence by amino acid sequencing. The DNA sequence data matches all of the available known peptide sequences. The mature polypeptide contains 300 amino acids having a combined molecular weight of 32,974 daltons. A putative signal peptide of 21 amino acids is proposed from DNA sequence data. The chloroperoxidase gene encodes three potential glycosylation sites recognized as Asn-X-Thr/Ser sequences. Three cysteine residues are found in the protein sequence. A small region around Cys87 bears a minimal homology to the active site of cytochrome P450cam. No other heme protein homologues can be detected. We propose that Cys87 serves as a thiolate ligand to the iron of heme prosthetic group. A rare arginine codon, AGG, is used three times out of twelve in contrast to the very infrequent use of this codon in E. coli or yeast.
PMID: 3774552 [PubMed - indexed for MEDLINE]
PMCID: PMC311834
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Cited by 5 PubMed Central articles
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Immobilization of Chloroperoxidase on Aminopropyl-Glass.
Kadima TA, Pickard MA.
Appl Environ Microbiol. 1990 Nov; 56(11):3473-3477.
[Appl Environ Microbiol. 1990]
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Replacement of the proximal heme thiolate ligand in chloroperoxidase with a histidine residue.
Yi X, Mroczko M, Manoj KM, Wang X, Hager LP.
Proc Natl Acad Sci U S A. 1999 Oct 26; 96(22):12412-7.
[Proc Natl Acad Sci U S A. 1999]
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Human thyroid peroxidase: complete cDNA and protein sequence, chromosome mapping, and identification of two alternately spliced mRNAs.
Kimura S, Kotani T, McBride OW, Umeki K, Hirai K, Nakayama T, Ohtaki S.
Proc Natl Acad Sci U S A. 1987 Aug; 84(16):5555-9.
[Proc Natl Acad Sci U S A. 1987]
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