title: Biochemical and proteomic analysis of the magnetosome membrane in Magnetospirillum gryphiswaldense.
abstract: We analyzed the biochemical composition of the magnetosome membrane (MM) in Magnetospirillum gryphiswaldense. Isolated magnetosomes were associated with phospholipids and fatty acids which were similar to phospholipids and fatty acids from other subcellular compartments (i.e., outer and cytoplasmic membranes) but were present in different proportions. The binding characteristics of MM-associated proteins were studied by selective solubilization and limited proteolysis. The MM-associated proteins were further analyzed by various proteomic approaches, including one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Edman and mass spectrometric (electrospray ionization-mass spectrometry-mass spectrometry) sequencing, as well as capillary liquid chromatography-mass spectrometry-mass spectrometry of total tryptic digests of the MM. At least 18 proteins were found to constitute the magnetosome subproteome, and most of these proteins are novel for M. gryphiswaldense. Except for MM22 and Mms16, all bona fide MM proteins (MMPs) were encoded by open reading frames in the mamAB, mamDC, and mms6 clusters in the previously identified putative magnetosome island. Eight of the MMPs display homology to known families, and some of them occur in the MM in multiple homologues. Ten of the MMPs have no known homologues in nonmagnetic organisms and thus represent novel, magnetotactic bacterium-specific protein families. Several MMPs display repetitive or highly acidic sequence patterns, which are known from other biomineralizing systems and thus may have relevance for magnetite formation.
authors.affiliation: Max-Planck-Institut für Marine Mikrobiologie, 28359 Bremen, Germany.
authors.forename: Karen
authors.initials: K
authors.lastname: Grünberg
authors.forename: Eva-Christina
authors.initials: EC
authors.lastname: Müller
authors.forename: Albrecht
authors.initials: A
authors.lastname: Otto
authors.forename: Regina
authors.initials: R
authors.lastname: Reszka
authors.forename: Dietmar
authors.initials: D
authors.lastname: Linder
authors.forename: Michael
authors.initials: M
authors.lastname: Kube
authors.forename: Richard
authors.initials: R
authors.lastname: Reinhardt
authors.forename: Dirk
authors.initials: D
authors.lastname: Schüler
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D005290:Ferric Compounds / Q000737:chemistry / Q000378:metabolism; D007425:Intracellular Membranes / Q000737:chemistry / Q000378:metabolism*; D042201:Magnetospirillum / Q000378:metabolism / Q000648:ultrastructure*; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D020543:Proteome
chemical_list: D001426:Bacterial Proteins; D005290:Ferric Compounds; D008565:Membrane Proteins; D020543:Proteome