title: Posttranslational modification and sequence variation of redox-active proteins correlate with biofilm life cycle in natural microbial communities.
abstract: Characterizing proteins recovered from natural microbial communities affords the opportunity to correlate protein expression and modification with environmental factors, including species composition and successional stage. Proteogenomic and biochemical studies of pellicle biofilms from subsurface acid mine drainage streams have shown abundant cytochromes from the dominant organism, Leptospirillum Group II. These cytochromes are proposed to be key proteins in aerobic Fe(II) oxidation, the dominant mode of cellular energy generation by the biofilms. In this study, we determined that posttranslational modification and expression of amino-acid sequence variants change as a function of biofilm maturation. For Cytochrome₅₇₉ (Cyt₅₇₉), the most abundant cytochrome in the biofilms, late developmental-stage biofilms differed from early-stage biofilms in N-terminal truncations and decreased redox potentials. Expression of sequence variants of two monoheme c-type cytochromes also depended on biofilm development. For Cyt(572), an abundant membrane-bound cytochrome, the expression of multiple sequence variants was observed in both early and late developmental-stage biofilms; however, redox potentials of Cyt₅₇₂ from these different sources did not vary significantly. These cytochrome analyses show a complex response of the Leptospirillum Group II electron transport chain to growth within a microbial community and illustrate the power of multiple proteomics techniques to define biochemistry in natural systems.
authors.affiliation: Physical and Life Sciences Directorate, Lawrence Livermore National Laboratory, Livermore, CA, USA. SWSinger@lbl.gov
authors.forename: Steven W
authors.initials: SW
authors.lastname: Singer
authors.forename: Brian K
authors.initials: BK
authors.lastname: Erickson
authors.forename: Nathan C
authors.initials: NC
authors.lastname: VerBerkmoes
authors.forename: Mona
authors.initials: M
authors.lastname: Hwang
authors.forename: Manesh B
authors.initials: MB
authors.lastname: Shah
authors.forename: Robert L
authors.initials: RL
authors.lastname: Hettich
authors.forename: Jillian F
authors.initials: JF
authors.lastname: Banfield
authors.forename: Michael P
authors.initials: MP
authors.lastname: Thelen
mesh_terms: D018407:Bacterial Physiological Phenomena; D018441:Biofilms / Q000254:growth & development*; D003580:Cytochromes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004783:Environmental Microbiology; D010084:Oxidation-Reduction; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000032:analysis; D017384:Sequence Deletion
chemical_list: D003580:Cytochromes; D020543:Proteome