Molecular cloning of complementary DNAs encoding two cationic peroxidases from cultivated peanut cells.
We have isolated, cloned, and characterized two cDNAs corresponding to the mRNAs for cationic peroxidases synthesized by cultured peanut cells. The first clone was obtained from a phage lambda gt11 library screened with antibodies directed against the major secreted isozyme. Its predicted amino acid sequence, deduced from the 1228-base-pair (bp) cDNA, revealed a 22-amino acid signal peptide and a 294-amino acid mature protein (Mr, 31,228). The second clone was isolated from a lambda gt10 library screened with oligonucleotides corresponding to the regions for acid/base catalysis and the fifth ligand of heme. This cDNA (1344 bp) encodes a protein (330 amino acids) with a mature peptide of 307 residues (Mr, 32,954). The two peanut peroxidases are 46% homologous. The estimated gene copy numbers of these peroxidases might be close to 1 or 2 per haploid genome. A comparison of the amino acid sequence of these peanut peroxidases with other known isozymes shows two already known regions of homology (the region for acid/base catalysis and the fifth ligand of heme). Moreover, some new characteristics appeared such as a glycosylation site identical in five of the seven isozymes, a putative antigenic determinant common to all the isozymes, and a region of the highest homology. A secondary structure prediction showed that it corresponds to a 16-amino acid helix linked to the next one by a long stretch of beta strands and coils and might represent a critical structural element.
Buffard
D
D
Institut des Sciences Végétales, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.
Breda
C
C
van Huystee
R B
RB
Asemota
O
O
Pierre
M
M
Ha
D B
DB
Esnault
R
R
D000595:Amino Acid Sequence; D010367:Arachis / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D015723:Gene Library; D005796:Genes; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D010544:Peroxidases / Q000235:genetics*
D009841:Oligonucleotides; D004247:DNA; D010544:Peroxidases