title: Performance of a novel sieving matrix of poly(vinyl alcohol)/acrylamide copolymer in electrophoretic separations of high molecular weight proteins from red cell membrane.
abstract: The analysis of high molecular weight (HMW) proteins from complex mixtures is still a challenge in proteomics. This work introduces a novel hydrogel obtained by the copolymerization of an allyl-PVA derivative with acrylamide and bisacrylamide and applies this matrix to the electrophoretic separation of HMW proteins. By inducing gelation of polyacrylamide in the presence of variable amounts of allyl-PVA, it is possible to control and vary the average gel porosity. This gel is easy to produce and handle and offers the advantage of being highly mechanically resistant and macroporous. The new matrix was tested in mono-dimensional separations of complex protein mixtures extracted from red cell membranes with different detergents. The improved performance of this macroporous matrix allowed to identify new proteins by MS and immunoblot analysis using specific antibodies. In particular, the resolution of proteins ranging in size between 97 and 279 kDa was greatly improved here compared to standard polyacrylamide gels, suggesting that this matrix can be a useful tool in routine analysis of HMW proteins in cell biology.
authors.affiliation: Department of Medicine, University of Verona, Verona, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Matte
authors.forename: Laura
authors.initials: L
authors.lastname: Sola
authors.forename: Marcella
authors.initials: M
authors.lastname: Chiari
authors.forename: Carlo
authors.initials: C
authors.lastname: Tomelleri
authors.forename: Roberto
authors.initials: R
authors.lastname: Consonni
authors.forename: Franco
authors.initials: F
authors.lastname: Turrini
authors.forename: Lucia De
authors.initials: LD
authors.lastname: Franceschi
mesh_terms: D020106:Acrylamide; D000180:Acrylic Resins / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004910:Erythrocyte Membrane / Q000737:chemistry*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification*; D008970:Molecular Weight; D011142:Polyvinyl Alcohol / Q000737:chemistry*; D016062:Porosity; D000099166:Polyacrylamides
chemical_list: D020106:Acrylamide; D000180:Acrylic Resins; D008565:Membrane Proteins; D011142:Polyvinyl Alcohol; D000099166:Polyacrylamides; C016680:polyacrylamide gels
keywords: High molecular weight proteins; Macroporous sieving matrix; Poly(vinyl alcohol); Polyacrylamide; Proteomics