title: Regulation of the catalytic behaviour of L-form starch phosphorylase from sweet potato roots by proteolysis.
abstract: Starch phosphorylase (SP) is an enzyme used for the reversible phosphorolysis of the alpha-glucan in plant cells. When compared to its isoform in an animal cell, glycogen phosphorylase, a peptide containing 78 amino acids (L78) is inserted in the centre of the low-affinity type starch phosphorylase (L-SP). We found that the amino acid sequence of L78 had several interesting features including the presence of a PEST region, which serves as a signal for rapid degradation. Indeed, most L-SP molecules isolated from mature sweet potato roots were nicked in the middle of a molecule, but still retained their tertiary or quaternary structures, as well as full catalytic activity. The nicking sites on the L78 were identified by amino acid sequencing of these peptides, which also enabled us to propose a proteolytic process for L-SP. Enzyme kinetic studies of L-SP in the direction of starch synthesis indicated that the Km decreased during the proteolytic process when starch was used as the limiting substrate, but the Km for the other substrate (Glc-1-P) increased. On the other hand, the maximum velocities (Vmax) increased for both substrates. Mobility of the nicked L-SP was retarded on a native polyacrylamide gel containing soluble starch, indicating the increased affinity for starch. Results in this study suggested that L78 and its proteolytic modifications might play a regulatory role on the catalytic behaviour of L-SP in starch biosynthesis.
authors.affiliation: Department of Agricultural Chemistry, National Taiwan University, Taipei, Taiwan 106.
authors.forename: Han-Min
authors.initials: HM
authors.lastname: Chen
authors.forename: Shih-Chung
authors.initials: SC
authors.lastname: Chang
authors.forename: Chi-Chen
authors.initials: CC
authors.lastname: Wu
authors.forename: Ting-Shen
authors.initials: TS
authors.lastname: Cuo
authors.forename: Jiann-Shing
authors.initials: JS
authors.lastname: Wu
authors.forename: Rong-Huay
authors.initials: RH
authors.lastname: Juang