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Enzymatic characterization of starch synthase III from kidney bean (Phaseolus vulgaris L.).
FEBS J. 2007 Sep; 274(17):4550-60.FJ

Abstract

In plants and green algae, several starch synthase isozymes are responsible for the elongation of glucan chains in the biosynthesis of amylose and amylopectin. Multiple starch synthase isozymes, which are classified into five major classes (granule-bound starch synthases, SSI, SSII, SSIII, and SSIV) according to their primary sequences, have distinct enzymatic properties. All the starch synthase isozymes consist of a transit peptide, an N-terminal noncatalytic region (N-domain), and a C-terminal catalytic region (C-domain). To elucidate the enzymatic properties of kidney bean (Phaseolus vulgaris L.) SSIII and the function of the N-domain of kidney bean SSIII, three recombinant proteins were constructed: putative mature recombinant SSIII, recombinant kidney bean SSIII N-domain, and recombinant kidney bean SSIII C-domain. Purified recombinant kidney bean SSIII displayed high specific activities for primers as compared to the other starch synthase isozymes from kidney bean. Kinetic analysis showed that the high specific activities of recombinant kidney bean SSIII are attributable to the high k(cat) values, and that the K(m) values of recombinant kidney bean SSIII C-domain for primers were much higher than those of recombinant kidney bean recombinant SSIII. Recombinant kidney bean SSIII and recombinant kidney bean SSIII C-domain had similar chain-length specificities for the extension of glucan chains, indicating that the N-domain of kidney bean SSIII does not affect the chain-length specificity. Affinity gel electrophoresis indicated that recombinant kidney bean SSIII and recombinant kidney bean SSIII N-domain have high affinities for amylose and amylopectin. The data presented in this study provide direct evidence for the function of the N-domain of kidney bean SSIII as a carbohydrate-binding module.

Authors+Show Affiliations

Senoura T
Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Sapporo, Japan.
Asao A
No affiliation info available
Takashima Y
No affiliation info available
Isono N
No affiliation info available
Hamada S
No affiliation info available
Ito H
No affiliation info available
Matsui H
No affiliation info available

MeSH

Catalytic DomainDNA, ComplementaryElectrophoresis, Polyacrylamide GelIsoenzymesMolecular Sequence DataPhaseolusPlants, Genetically ModifiedRecombinant ProteinsReverse Transcriptase Polymerase Chain ReactionStarch Synthase

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17681016

Citation

Senoura, Takeshi, et al. "Enzymatic Characterization of Starch Synthase III From Kidney Bean (Phaseolus Vulgaris L.)." The FEBS Journal, vol. 274, no. 17, 2007, pp. 4550-60.
Senoura T, Asao A, Takashima Y, et al. Enzymatic characterization of starch synthase III from kidney bean (Phaseolus vulgaris L.). FEBS J. 2007;274(17):4550-60.
Senoura, T., Asao, A., Takashima, Y., Isono, N., Hamada, S., Ito, H., & Matsui, H. (2007). Enzymatic characterization of starch synthase III from kidney bean (Phaseolus vulgaris L.). The FEBS Journal, 274(17), 4550-60.
Senoura T, et al. Enzymatic Characterization of Starch Synthase III From Kidney Bean (Phaseolus Vulgaris L.). FEBS J. 2007;274(17):4550-60. PubMed PMID: 17681016.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enzymatic characterization of starch synthase III from kidney bean (Phaseolus vulgaris L.). AU - Senoura,Takeshi, AU - Asao,Ayako, AU - Takashima,Yoshinori, AU - Isono,Naoto, AU - Hamada,Shigeki, AU - Ito,Hiroyuki, AU - Matsui,Hirokazu, Y1 - 2007/08/06/ PY - 2007/8/8/pubmed PY - 2007/10/27/medline PY - 2007/8/8/entrez SP - 4550 EP - 60 JF - The FEBS journal JO - FEBS J VL - 274 IS - 17 N2 - In plants and green algae, several starch synthase isozymes are responsible for the elongation of glucan chains in the biosynthesis of amylose and amylopectin. Multiple starch synthase isozymes, which are classified into five major classes (granule-bound starch synthases, SSI, SSII, SSIII, and SSIV) according to their primary sequences, have distinct enzymatic properties. All the starch synthase isozymes consist of a transit peptide, an N-terminal noncatalytic region (N-domain), and a C-terminal catalytic region (C-domain). To elucidate the enzymatic properties of kidney bean (Phaseolus vulgaris L.) SSIII and the function of the N-domain of kidney bean SSIII, three recombinant proteins were constructed: putative mature recombinant SSIII, recombinant kidney bean SSIII N-domain, and recombinant kidney bean SSIII C-domain. Purified recombinant kidney bean SSIII displayed high specific activities for primers as compared to the other starch synthase isozymes from kidney bean. Kinetic analysis showed that the high specific activities of recombinant kidney bean SSIII are attributable to the high k(cat) values, and that the K(m) values of recombinant kidney bean SSIII C-domain for primers were much higher than those of recombinant kidney bean recombinant SSIII. Recombinant kidney bean SSIII and recombinant kidney bean SSIII C-domain had similar chain-length specificities for the extension of glucan chains, indicating that the N-domain of kidney bean SSIII does not affect the chain-length specificity. Affinity gel electrophoresis indicated that recombinant kidney bean SSIII and recombinant kidney bean SSIII N-domain have high affinities for amylose and amylopectin. The data presented in this study provide direct evidence for the function of the N-domain of kidney bean SSIII as a carbohydrate-binding module. SN - 1742-464X UR - https://www.unboundmedicine.com/medline/citation/17681016/Enzymatic_characterization_of_starch_synthase_III_from_kidney_bean__Phaseolus_vulgaris_L___ DB - PRIME DP - Unbound Medicine ER -