title: Limited proteolysis of chloroplast glyceraldehyde-3-phosphate dehydrogenase (NADP) from Spinacia oleracea.
abstract: The structural and functional properties of chloroplast glyceraldehyde-3-P-dehydrogenase I (D-Glyceraldehyde-3-phosphate: NADP oxidoreductase (phosphorylating) EC 1.2.1.13) from Spinacia oleracea were investigated by limited proteolysis. The enzyme is insensitive to trypsin and chymotrypsin, while Staphylococcus aureus V8 protease cleaves the C-terminal region of its subunits. Subunit A (36 kDa) is only partially cleaved at Glu 317. No intact subunit B (39 kDa) is found at the end of the proteolytic experiment: two forms are originated from this subunit which is cleaved at Glu 342 and Glu 320. Proteolytic cleavage at these sites does not significantly alter enzymatic activity, but leads to destabilization of the protein. Unlike the intact parent enzyme (600 kDa) the cleaved enzyme behaves as a 150-kDa species in size exclusion chromatography.
authors.affiliation: Dipartimento di Biochimica, Università di Pavia, Italia.
authors.forename: M C
authors.initials: MC
authors.lastname: Zapponi
authors.forename: P
authors.initials: P
authors.lastname: Iadarola
authors.forename: M
authors.initials: M
authors.lastname: Stoppini
authors.forename: G
authors.initials: G
authors.lastname: Ferri
mesh_terms: D000445:Aldehyde Oxidoreductases / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D002736:Chloroplasts / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005973:Glutamine / Q000737:chemistry; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010766:Phosphorylation; D017421:Sequence Analysis; D012697:Serine Endopeptidases / Q000378:metabolism; D014675:Vegetables / Q000201:enzymology
chemical_list: D005973:Glutamine; D000445:Aldehyde Oxidoreductases; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase