title: Synthesis and resistance to in vitro proteolysis of transglutaminase cross-linked phaseolin, the major storage protein from Phaseolus vulgaris.
abstract: The ability of phaseolin to act as an acyl donor and acceptor substrate of transglutaminase was studied by using an enzyme isolated from Streptoverticillium mobarense. Phaseolin, a trimeric storage protein from Phaseolus vulgaris L., was shown to possess both glutamine and lysine residues reactive for the enzyme. The extent of transglutaminase-catalyzed cross-linking has been studied in function of both incubation time and enzyme concentration. Native- and SDS-PAGE demonstrated that phaseolin is intra- and intermolecularly cross-linked by transglutaminase and gives rise to different polymers as well as to modified forms of the protein having a similar molecular weight but lower Stokes radius if compared to unmodified phaseolin. Cross-linked phaseolin was found to be more resistant to proteolytic cleavage than the unmodified counterpart, as demonstrated by in vitro trypsin and pepsin digestion experiments. This behavior could suggest novel possible uses of the transglutaminase-modified phaseolin.
authors.affiliation: Department of Food Science and School of Biotechnological Sciences, University of Naples Federico II, Parco Gussone, 80055 Portici, Naples, Italy. loredana.mariniello@unina.it
authors.forename: Loredana
authors.initials: L
authors.lastname: Mariniello
authors.forename: Concetta Valeria L
authors.initials: CV
authors.lastname: Giosafatto
authors.forename: Prospero
authors.initials: P
authors.lastname: Di Pierro
authors.forename: Angela
authors.initials: A
authors.lastname: Sorrentino
authors.forename: Raffaele
authors.initials: R
authors.lastname: Porta
mesh_terms: D003198:Computer Simulation; D003432:Cross-Linking Reagents / Q000737:chemistry; D004355:Drug Stability; D004795:Enzyme Stability; D007700:Kinetics; D010447:Peptide Hydrolases; D027805:Phaseolus / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism*; D011503:Transglutaminases / Q000378:metabolism*
chemical_list: D003432:Cross-Linking Reagents; D010940:Plant Proteins; C048826:phaseolin protein, Phaseolus vulgaris; D011503:Transglutaminases; D010447:Peptide Hydrolases