title: Purification of plant complex protein extracts in non-denaturing conditions by in-solution isoelectric focusing.
abstract: An alternative approach for plant complex protein extracts pre-purification by in-solution isoelectric focusing in non-denaturing conditions is presented. The separation of biologically active proteins, in narrow ranges of isoelectric point (pI) was obtained by a modified OFFGEL electrophoresis. Two different water-soluble protein extracts from Phragmites leaves were fractionated into 24 fractions within a 3-10 pI range at 10 °C in the absence of denaturing/reducing agents. One-dimensional electrophoretic analysis revealed different protein distribution patterns and the effective fractionation of both protein extracts. Peroxidase activity of each fraction confirmed that proteins remained active and pre-purification occurred. Biological triplicates assured the needed reproducibility.
authors.affiliation: Centre for Natural Resources and the Environment (CERENA), Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal.
authors.forename: R A
authors.initials: RA
authors.lastname: Ferreira
authors.affiliation: Centre for Natural Resources and the Environment (CERENA), Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal. Electronic address: susetedias@tecnico.ulisboa.pt.
authors.forename: S
authors.initials: S
authors.lastname: Martins-Dias
mesh_terms: D007525:Isoelectric Focusing / Q000379:methods; D010936:Plant Extracts / Q000737:chemistry*; D018515:Plant Leaves / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D006109:Poaceae / Q000737:chemistry*
chemical_list: D010936:Plant Extracts; D010940:Plant Proteins
keywords: In-solution IEF; Non-denaturing conditions; OFFGEL electrophoresis; Phragmites sp.; Plant protein purification; pI