Identification of the Methanococcus voltae S-layer structural gene

Konisky, J; Lynn, D. G.; Hoppert, Michael; Mayer, F; Haney, Paul

doi:10.1128/jb.176.6.1790-1792.1994

Cited by 20 publications

(11 citation statements)

References 13 publications

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“…In M. voltae, the only other protein with a proposed leader peptide is the S-layer protein, which was initially reported to possess a 12-amino-acid long leader peptide (Konisky et al 1994). However, recent comparative evidence suggests that a 28-amino-acid leader peptide similar to canonical leader peptides may in fact be present.…”

Section: Resultsmentioning

confidence: 98%

Identification of amino acids in the leader peptide of Methanococcus voltae preflagellin that are important in posttranslational processing

Thomas

Chao

Jarrell

2001

Archives of Microbiology

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Archaeal flagellins are made initially as preproteins with short, positively charged leader peptides. Analysis of all available archaeal preflagellin sequences indicates that the -1 position is always held by a glycine while the -2 and -3 positions are almost always held by charged amino acids. To evaluate the importance of these and other amino acids in the leader peptides of archaeal flagellins for processing by a peptidase, Methanococcus voltae mutant FlaB2 preflagellin genes were generated by PCR and the proteins tested in a methanogen preflagellin peptidase assay that detects the removal of the leader peptide from preflagellin. When the -1 position was changed from glycine to other amino acids tested, no cleavage was observed by the peptidase, with the exception of a change to alanine at which poor, partial processing was observed. Amino acid substitutions at the -2 lysine position resulted in a complete loss of processing by the peptidase, while changes at the -3 lysine resulted in partial processing. A mutant preflagellin with a leader peptide shortened from 12 amino acids to 6 amino acids was not processed. When the invariant glycine residue present at position +3 was changed to a valine, no processing of this mutant preflagellin was observed. The identification of critical amino acids in FlaB2 required for proper processing suggests that a specific preflagellin peptidase may cleave archaeal flagellins by recognition of a conserved sequence of amino acids.

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Section: Resultsmentioning

confidence: 98%

Identification of amino acids in the leader peptide of Methanococcus voltae preflagellin that are important in posttranslational processing

Thomas

Chao

Jarrell

2001

Archives of Microbiology

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“…Glycoproteins MM1976, MA0829, and Methanococcus voltae S-layer protein Q5083340 contain tandem duplicated domains of ∼250 amino acids which, in the Methanosarcinaceae , are referenced variously as PF007752, IPR006457 or DUF1608. However, BlastP comparisons find no significant similarity between the Methanococcus and Methanosarcinaceae proteins.…”

Section: Discussionmentioning

confidence: 99%

S-layer, Surface-Accessible, and Concanavalin A Binding Proteins of Methanosarcina acetivorans and Methanosarcina mazei

et al. 2009

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The outermost cell envelope structure of many archaea and bacteria contains a proteinaceous lattice termed the surface layer or S-layer. It is typically composed of only one or two abundant, often posttranslationally modified proteins that self-assemble to form the highly organized arrays. Surprisingly, over a hundred proteins were annotated to be S-layer components in the archaeal species Methanosarcina acetivorans C2A and Methanosarcina mazei Gö1, reflecting limitations of current predictions. An in vivo biotinylation methodology was devised to affinity tag surface-exposed proteins while overcoming unique challenges in working with these fragile organisms. Cells were adapted to growth under N 2 fixing conditions, thus minimizing free amines reactive to the NHSlabel, and high pH media compatible with the acylation chemistry was used. A 3-phase separation procedure was employed to isolate intact, labeled cells from lysed-cell derived proteins. Streptavidin affinity enrichment followed by stringent wash conditions removed non-specifically bound proteins. This methodology revealed S-layer proteins in M. acetivorans C2A and M. mazei Gö1 to be MA0829 and MM1976, respectively. Each was demonstrated to exist as multiple glycosylated forms using SDS-PAGE coupled with glycoprotein-specific staining, and by interaction with the lectin, Concanavalin A. A number of additional surface-exposed proteins and glycoproteins were identified and included all three subunits of the thermosome: the latter suggests that the chaperonin complex is both surface-and cytoplasmically-localized. This approach provides an alternative strategy to study surface proteins in the archaea.

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“…E SDS-PAGE (10% acrylamide) with an enzyme preparation that was obtained by using buffers lacking KCl of an S-layer structural protein (MJ0822; accession number U67526) (Bult et al 1996); the deduced molecular mass for this polypeptide was 60.5 kDa. A discrepancy between the SDS-PAGE-derived and nucleic-acid-sequence-deduced molecular masses has also been found for the Methanococcus voltae S-layer protein (Konisky et al 1994) and could be an indication of protein glycosylation or some other modification. The NH 2 -terminal sequence of the 110-kDa polypeptide corresponded to an ORF (MJ1156; accession number U67557) (Bult et al 1996) that has been annotated as a putative cell division control protein 48 (cdc48), AAA family (ATPase associated with diverse cellular activities).…”

Section: Primary Structures Identities For the Polypeptides In The Pymentioning

confidence: 93%

A stable archaeal pyruvate carboxylase from the hyperthermophile Methanococcus jannaschii

Mukhopadhyay

Patel

Wolfe

2000

Archives of Microbiology

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The pyruvate carboxylase (PYC) of the hyperthermophilic, strictly hydrogenotrophic, autotrophic and marine methanarchaeon Methanococcus jannaschii was purified to homogeneity. Optimal activity was at pH 8.5, > or = 80 degrees C, and a KCl concentration of 0.175 M. This enzyme is the most thermophilic PYC so far studied. Unlike the Methanobacterium thermoautotrophicum enzyme, Mc. jannaschii PYC was expressed in cells grown without an external source of biotin and in the purified form was stable during storage at 4, -20 and -80 degrees C. However, it was rapidly inactivated at 80 degrees C. The enzyme was insensitive to aspartate and glutamate, mildly inhibited by alpha-ketoglutarate, and was strongly inhibited by ATP and ADP (apparent Km, for ATP, 0.374 +/- 0.039 mM; apparent Ki for ATP, 5.34 +/- 2.14 mM; Ki for ADP, 0.89 +/- 0.18 mM). It was also strongly inhibited when the Mg2+ concentration in the assay exceeded that of ATP. Thus, this stable PYC could serve as a model for mechanistic studies on archaeal PYCs. It was apparently an alpha4beta4-type PYC composed of a non-biotinylated 55.5-kDa subunit (PYCA) and a 64.2-kDa biotinylated subunit (PYCB). The determined NH2-terminal sequences for these subunits provided additional support for our earlier proposal to rename the ORFs MJ1229 and MJ1231 in the NCBI Mc. jannaschii genome sequence database as PYCA and PYCB, respectively; even very recently, these have been misidentified as a subunit of acetyl-CoA carbxoylase (AccC) and the alpha-subunit of ion-pumping oxaloacetate decarboxylase (OADalpha), respectively.

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Identification of the Methanococcus voltae S-layer structural gene

Cited by 20 publications

References 13 publications

Identification of amino acids in the leader peptide of Methanococcus voltae preflagellin that are important in posttranslational processing

Identification of amino acids in the leader peptide of Methanococcus voltae preflagellin that are important in posttranslational processing

S-layer, Surface-Accessible, and Concanavalin A Binding Proteins of Methanosarcina acetivorans and Methanosarcina mazei

A stable archaeal pyruvate carboxylase from the hyperthermophile Methanococcus jannaschii

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