A cysteine endopeptidase isolated from castor bean endosperm microbodies processes the glyoxysomal malate dehydrogenase precursor protein

• Published in: Plant physiology (Bethesda) Vol. 113; no. 3; pp. 863 - 871
• Main Authors: Gietl, C. (Technical University of Munich, Munich, Germany.), Wimmer, B, Adamex, J, Kalousek, F
• Format: Journal Article
• Language: English
• Published: Rockville, MD American Society of Plant Physiologists 01.03.1997
• Subjects: ACILTRANSFERASA; ACTIVIDAD ENZIMATICA; ACTIVITE ENZYMATIQUE; ACYLTRANSFERASE; Amino Acid Sequence; Amino acids; ARGININA; ARGININE; Biological and medical sciences; BRASSICA NAPUS; CISTEINA; COMPOSICION QUIMICA; COMPOSITION CHIMIQUE; Cotyledons; CYSTEINE; Cysteine Endopeptidases - isolation & purification; Cysteine Endopeptidases - metabolism; Dehydrogenases; Development and Growth Regulation; Endosperm; ENDOSPERMA; ENDOSPERME; Enzyme Precursors - metabolism; Enzymes; Fabaceae - enzymology; Fundamental and applied biological sciences. Psychology; GERMINACION; GERMINATION; Hydrolysis; Malate Dehydrogenase - metabolism; MALATE DESHYDROGENASE; MALATO DESHIDROGENASA; MEDIO DE CULTIVO; Metabolism; Microbodies; Microbodies - enzymology; MILIEU DE CULTURE; Molecular Sequence Data; ORGANITE CELLULAIRE; ORGANULOS CITOPLASMICOS; PEPTIDASAS; PEPTIDASE; Peroxisomes; PHASEOLUS VULGARIS; Plant physiology and development; Plants, Medicinal; Protein precursors; Protein Processing, Post-Translational; PROTEINAS; PROTEINE; Proteins; PURIFICACION; PURIFICATION; RAT; RATA; RICINUS COMMUNIS; Seedlings; Substrate Specificity; Trypsin - metabolism; VIGNA MUNGO; Purification; Enzyme; Cysteine endopeptidases; Homology; Malate dehydrogenase; Microbody; Characterization; Ripening; Peptidases; Glyoxysome; Dicotyledones; Angiospermae; N terminal-Sequence; Euphorbiaceae; Hydrolases; Spermatophyta; Endosperm; Oxidoreductases; Aminoacid sequence; Ricinus communis
• ISSN: 0032-0889; 1532-2548
• DOI: 10.1104/pp.113.3.863

A plant cysteine endopeptidase with a molecular mass of 35 kD was purified from microbodies of germinating castor bean (Ricinus communis) endosperm by virtue of its capacity to specifically process the glyoxysomal malate dehydrogenase precursor protein to the mature subunit in vitro. Processing of the glyoxysomal malate dehydrogenase precursor occurs sequentially in three steps, the first intermediate resulting from cleavage after arginine-13 within the presequence and the second from cleavage after arginine-33. The endopeptidase is unable to remove the presequences of prethiolases from rape (Brassica napus) glyoxysomes and rat peroxisomes at the expected cleavage site. Protein sequence analysis of N-terminal and internal peptides revealed high identity to the mature papain-type cysteine endopeptidases from cotyledons of germinating mung bean (Vigna mungo) and French bean (Phaseolus vulgaris) seeds. These endopeptidases are synthesized with an extended pre-/prosequence at the N terminus and have been considered to be processed in the endoplasmic reticulum and targeted to protein-storing vacuoles