Pro domain peptide of HGCP-Iv cysteine proteinase inhibits nematode cysteine proteinases.
Cysteine proteinases (CPs) are synthesized as zymogens and converted to mature proteinase forms by proteolytic cleavage and release of their pro domain peptides. A cDNA encoding a papain-like CP, called hgcp-Iv, was isolated from a Heterodera glycines J2 cDNA library, expressed and utilized to assess the ability of its propeptide to inhibit proteinase in its active form. The hgcp-Iv cDNA sequence encodes a polypeptide of 374 amino acids with the same domain organization as other cathepsin L-like CPs, including a hydrophobic signal sequence and a pro domain region. HGCP-Iv, produced in Escherichia coli as a fusion protein with thioredoxin, degrades the synthetic peptide benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin and is inhibited by E-64, a substrate and inhibitor commonly used for functional characterization of CPs. Recombinant propeptides of HGCP-Iv, expressed in E. coli, presented high inhibitory activity in vitro towards its cognate enzyme and proteinase activity of Meloidogyne incognita females, suggesting its usefulness in inhibiting nematode CPs in biological systems. Cysteine proteinases from other species produced no noticeable activity.
Silva
Francine B
FB
Departamento de Biologia Celular, Universidade de Brasília, Brasília, DF, Brasil.
Batista
João A N
JA
Marra
Brener M
BM
Fragoso
Rodrigo R
RR
Monteiro
Ana Carolina S
AC
Figueira
Edson L Z
EL
Grossi-de-Sá
Maria Fátima
MF
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003546:Cysteine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics; D005260:Female; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics* / Q000378:metabolism; D010935:Plant Diseases / Q000469:parasitology*; D016133:Polymerase Chain Reaction; D014431:Tylenchoidea / Q000201:enzymology* / Q000235:genetics
D015853:Cysteine Proteinase Inhibitors; D018076:DNA, Complementary; D018509:DNA, Helminth; D010455:Peptides; D003546:Cysteine Endopeptidases