Characterization of the wound‐induced metallocarboxypeptidase inhibitor from potato 1

FEBS Letters

Avilés, Francesc X²  Querol, Enrique²  Ludevid, Dolors¹  Prat, Salomé¹  Villanueva, Josep²  Canals, Francesc² 

[1] Departament de Genètica Molecular, Centre de Investigació i Desenvolupament, CSIC, Jordi Girona 18–26, 08034 Barcelona, Spain;Institut de Biologia Fonamental and Departament de Bioquı́mica, Universitat Autònoma de Barcelona, 08193 Bellaterra (Barcelona), Spain

关键词: Metallocarboxypeptidase inhibitor;    Precursor protein;    Vacuolar sorting;    Wound-inducible expression;    (Solanum tuberosum);    ABA;    abscisic acid;    MCPI;    metallocarboxypeptidase inhibitor;    PCI;    potato carboxypeptidase inhibitor;    CPA;    carboxypeptidase A;    CPB;    carboxypeptidase B;    JA;    jasmonic acid;    proPCI-Ct;    carboxy-terminal proPCI precursor;    Ct;    carboxy-terminal;    MALDI-TOF;    matrix-assisted laser desorption ionization time of flight;   

DOI  :  10.1016/S0014-5793(98)01447-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A partial cDNA clone for the potato wound-inducible metallocarboxypeptidase inhibitor (PCI) was isolated from a cDNA library constructed from mRNA of abscisic acid (ABA)-treated potato leaves. The full 5′ region of the cDNA was obtained through a RACE-PCR protocol. PCI mRNA encodes a precursor polypeptide which comprises a 29 residue N-terminal signal peptide, a 27 residue N-terminal pro-region, the 39 residue mature PCI protein, and a 7 residue C-terminal extension. Northern blot analysis demonstrates that the PCI gene is transcriptionally activated by wounding, and wound signaling can be induced by ABA and jasmonic acid. Subcellular localization of the protein was investigated by immunocytochemistry and electron microscopy, showing that PCI accumulates within the vacuole. A partial PCI precursor form, comprising the mature protein and the C-terminal extension, has been expressed in Escherichia coli and characterized. Its inability to inhibit carboxypeptidases, and stability to carboxypeptidase digestion, suggest that the C-terminal pro-domain may have, besides a probable vacuolar sorting function, a role in modulation of the inhibitory activity of PCI.

【 授权许可】

Unknown