title: Phenotypic subunit composition of the tobacco (Nicotiana tabacum L.) vacuolar-type H(+)-translocating ATPase.
abstract: The model plant tobacco (Nicotiana tabacum L.) was chosen for a survey of the subunit composition of the V-ATPase at the protein level. V-ATPase was purified from tobacco leaf cell tonoplasts by solubilization with the nonionic detergent Triton X-100 and immunoprecipitation. In the purified fraction 12 proteins were present. By matrix-assisted laser-desorption ionization mass spectrometry (MALDI-MS) and amino acid sequencing 11 of these polypeptides could be identified as subunits A, B, C, D, F, G, c, d and three different isoforms of subunit E. The polypeptide which could not be identified by MALDI analysis might represent subunit H. The data presented here, for the first time, enable an unequivocal identification of V-ATPase subunits after gel electrophoresis and open the possibility to assign changes in polypeptide composition to variations in respective V-ATPase subunits occurring as a response to environmental conditions or during plant development.
authors.affiliation: Darmstadt University of Technology, Institute of Botany, Schnittspahnstrasse 3-5, D-64287, Darmstadt, Germany.
authors.forename: Martina
authors.initials: M
authors.lastname: Drobny
authors.forename: Martina
authors.initials: M
authors.lastname: Schnölzer
authors.forename: Sabine
authors.initials: S
authors.lastname: Fiedler
authors.forename: Ulrich
authors.initials: U
authors.lastname: Lüttge
authors.forename: Elke
authors.initials: E
authors.lastname: Fischer-Schliebs
authors.forename: Anna-Luise
authors.initials: AL
authors.lastname: Christian
authors.forename: Rafael
authors.initials: R
authors.lastname: Ratajczak
mesh_terms: D000595:Amino Acid Sequence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D010641:Phenotype; D021122:Protein Subunits; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014026:Nicotiana / Q000201:enzymology* / Q000235:genetics; D014357:Trypsin; D025262:Vacuolar Proton-Translocating ATPases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D010446:Peptide Fragments; D021122:Protein Subunits; D014357:Trypsin; D025262:Vacuolar Proton-Translocating ATPases