title: Seryl-phosphorylation of soybean nodule sucrose synthase (nodulin-100) by a Ca2+-dependent protein kinase.
abstract: Sucrose synthase (SS; EC 2.4.1.13) was radiolabeled in situ by incubating detached soybean nodules with 32Pi. Phosphoamino acid analysis indicated that SS was phosphorylated on a serine residue(s). In-vitro phosphorylation of purified nodule SS by desalted nodule extracts was Ca2+-dependent. This SS-kinase was partially purified (approximately 2200-fold) from nodules harvested from illuminated plants. The molecular mass of the SS-kinase was about 55,000 on a Superdex 75 size-exclusion column or in a denaturing autophosphorylation gel. With either purified nodule SS or Syntide 2 as substrate, exogenous calmodulin and phosphatidylserine showed little or no effect on the in-vitro activity of this partially purified protein kinase. However, its activity was inhibited by W-7. The purified nodule SS-kinase (or CDPK) phosphorylated nodule PEP carboxylase (PEPC; EC 4.1.1.31) in the presence of Ca2+. In contrast, a partially purified nodule PEPC-kinase preparation was incapable of phosphorylating nodule SS. Unlike nodule PEPC [Zhang et al. (1995) Plant Physiol. 108, 1561-1568], the phosphorylation state of SS is not likely modulated in planta by photosynthate supply from the shoots.
authors.affiliation: Department of Biochemistry, University of Nebraska-Lincoln, George W. Beadle Center, 68588-0664, USA.
authors.forename: X Q
authors.initials: XQ
authors.lastname: Zhang
authors.forename: R
authors.initials: R
authors.lastname: Chollet
mesh_terms: D005964:Glucosyltransferases / Q000737:chemistry / Q000378:metabolism*; D008565:Membrane Proteins; D010766:Phosphorylation; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D012694:Serine / Q000378:metabolism; D013025:Glycine max / Q000201:enzymology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases
chemical_list: D005964:Glucosyltransferases; D008565:Membrane Proteins; D010940:Plant Proteins; D011494:Protein Kinases; D012694:Serine; D017871:Calcium-Calmodulin-Dependent Protein Kinases; C045625:nodulin; C022699:sucrose synthase