title: Collagenase as a useful tool for the analysis of plant cellular peripheries.
abstract: A technique for the selective loosening of the cell wall structure and the isolation of proteins permanently knotted in the cell walls was elaborated. Following treatment with collagenase, some proteins, such as calreticulin (CRT) and auxin binding protein 1 (ABP1) were released from purified cell walls, most probably through destruction of respective interacting proteins. The results were confirmed by the immunolocalization of the ABP1 and CRT with confocal and electron microscopy. On the other hand, potential substrates of collagenase, among them annexin 1 have been recognized. Mass spectra of annexin 1 obtained after collagenase digestion and results from analysis of potential cleavage sites suggested that the mechanism of enzyme cleavage might not depend on the amino acid sequence. Summarizing, collagenase was found to be a very useful tool for exploring molecules involved in the functioning of cellular peripheries.
authors.affiliation: Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznań, Poland. Electronic address: magdalu@ibch.poznan.pl.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Luczak
authors.affiliation: Department of Plant Biotechnology, Jagiellonian University, Kraków, Poland.
authors.forename: Weronika
authors.initials: W
authors.lastname: Krzeszowiec-Jeleń
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warszawa, Poland.
authors.forename: Dorota
authors.initials: D
authors.lastname: Konopka-Postupolska
authors.affiliation: Department of Molecular and Cellular Biology, Adam Mickiewicz University, Poznań, Poland. Electronic address: przemow@amu.edu.pl.
authors.forename: Przemysław
authors.initials: P
authors.lastname: Wojtaszek
mesh_terms: D000595:Amino Acid Sequence; D002473:Cell Wall / Q000378:metabolism*; D017364:Collagenases / Q000378:metabolism*; D055767:Immobilized Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D059828:Plant Cells / Q000378:metabolism*; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D059748:Proteolysis
chemical_list: D055767:Immobilized Proteins; D010940:Plant Proteins; D017364:Collagenases
keywords: Annexin 1; Auxin-binding protein 1; Calreticulin; Cell wall; Collagenase