Browse
Submit Data
Databases
API
Help

Dataset Information

21 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Crystallization and biochemical characterization of an archaeal lectin from Methanococcus voltae A3.

ABSTRACT: A lectin from Methanococcus voltae A3 has been cloned, expressed, purified and characterized. The lectin appears to be specific for complex sugars. The protein crystallized in a tetragonal space group, with around 16 subunits in the asymmetric unit. Sequence comparisons indicate the lectin to have a ?-prism I fold, with poor homology to lectins of known three-dimensional structure.

SUBMITTER: Sivaji N

PROVIDER: S-EPMC5417321 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature

ACCESS DATA

Json Xml

Publications

Crystallization and biochemical characterization of an archaeal lectin from Methanococcus voltae A3.

Sivaji N N Abhinav K V KV Vijayan M M

Acta crystallographica. Section F, Structural biology communications 20170428 Pt 5

A lectin from Methanococcus voltae A3 has been cloned, expressed, purified and characterized. The lectin appears to be specific for complex sugars. The protein crystallized in a tetragonal space group, with around 16 subunits in the asymmetric unit. Sequence comparisons indicate the lectin to have a β-prism I fold, with poor homology to lectins of known three-dimensional structure. ...[more]

PMID: 28471363

Similar Datasets

Cloning and characterization of archaeal type I signal peptidase from Methanococcus voltae.

Project description:Archaeal protein trafficking is a poorly characterized process. While putative type I signal peptidase genes have been identified in sequenced genomes for many archaea, no biochemical data have been presented to confirm that the gene product possesses signal peptidase activity. In this study, the putative type I signal peptidase gene in Methanococcus voltae was cloned and overexpressed in Escherichia coli, the membranes of which were used as the enzyme source in an in vitro peptidase assay. A truncated, His-tagged form of the M. voltae S-layer protein was generated for use as the substrate to monitor the signal peptidase activity. With M. voltae membranes as the enzyme source, signal peptidase activity in vitro was optimal between 30 and 40 degrees C; it was dependent on a low concentration of KCl or NaCl but was effective over a broad concentration range up to 1 M. Processing of the M. voltae S-layer protein at the predicted cleavage site (confirmed by N-terminal sequencing) was demonstrated with the overexpressed archaeal gene product. Although E. coli signal peptidase was able to correctly process the signal peptide during overexpression of the M. voltae S-layer protein in vivo, the contribution of the E. coli signal peptidase to cleavage of the substrate in the in vitro assay was minimal since E. coli membranes alone did not show significant activity towards the S-layer substrate in in vitro assays. In addition, when the peptidase assays were performed in 1 M NaCl (a previously reported inhibitory condition for E. coli signal peptidase I), efficient processing of the substrate was observed only when the E. coli membranes contained overexpressed M. voltae signal peptidase. This is the first proof of expressed type I signal peptidase activity from a specific archaeal gene product.

| S-EPMC225044 | biostudies-literature

Methanococcus voltae A3 strain:A3

Project description:Methanogenic archaeon isolated from a salt-marsh

| PRJNA20131 | ENA

Methanococcus voltae

Project description:Methane-producing archaeon

| PRJNA13632 | ENA

Methanococcus voltae

Project description:Methanococcus voltae

| PRJNA20139 | ENA

Structure of a hexameric form of RadA recombinase from Methanococcus voltae.

Project description:Archaeal RadA proteins are close homologues of eukaryal Rad51 and DMC1 proteins and are remote homologues of bacterial RecA proteins. For the repair of double-stranded breaks in DNA, these recombinases promote a pivotal strand-exchange reaction between homologous single-stranded and double-stranded DNA substrates. This DNA-repair function also plays a key role in the resistance of cancer cells to chemotherapy and radiotherapy and in the resistance of bacterial cells to antibiotics. A hexameric form of a truncated Methanococcus voltae RadA protein devoid of its small N-terminal domain has been crystallized. The RadA hexamers further assemble into two-ringed assemblies. Similar assemblies can be observed in the crystals of Pyrococcus furiosus RadA and Homo sapiens DMC1. In all of these two-ringed assemblies the DNA-interacting L1 region of each protomer points inward towards the centre, creating a highly positively charged locus. The electrostatic characteristics of the central channels can be utilized in the design of novel recombinase inhibitors.

| S-EPMC3374503 | biostudies-literature

Isolation and characterization of the 7S RNA gene from Methanococcus voltae.

Project description:The gene encoding the 7S RNA of the archaebacterium Methanococcus voltae has been isolated. The gene occurs as a single copy within the genome and encodes an RNA molecule approximately 300 nucleotides in length. The M. voltae RNA molecule exhibits a strong similarity to both archaebacterial and eucaryotic 7S RNAs in terms of overall secondary structure, while the primary sequence is conserved to a lesser degree. All 7S RNA molecules possess a specific structural domain which is highly conserved in terms of both primary sequence and secondary structure, possibly representing a functional site of the molecule. Conservation of the 7S RNA molecule suggests that it is the descendant of a subcellular structure present before the divergence of the archaebacterial and eucaryotic kingdoms. The M. voltae 7S RNA gene is flanked both 5' and 3' by regions of extremely A + T-rich DNA. The 5'-flanking region contains several potential promoter sequences for archaebacterial RNA polymerases. One such sequence occurs as three direct repeats and bears a strong similarity to sequences found upstream of other archaebacterial genes. The 3'-flanking region contains a strong signal for the termination of transcription.

| S-EPMC210199 | biostudies-other

Identification and localization of flagellins FlaA and FlaB3 within flagella of Methanococcus voltae.

Project description:Methanococcus voltae possesses four flagellin genes, two of which (flaB1 and flaB2) have previously been reported to encode major components of the flagellar filament. The remaining two flagellin genes, flaA and flaB3, are transcribed at lower levels, and the corresponding proteins remained undetected prior to this work. Electron microscopy examination of flagella isolated by detergent extraction of whole cells revealed a curved, hook-like region of varying length at the end of a long filament. Enrichment of the curved region of the flagella resulted in the identification of FlaB3 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and N-terminal sequencing, and the localization of this flagellin to the cell-proximal portion of the flagellum was confirmed through immunoblotting and immunoelectron microscopy with FlaB3-specific antibodies, indicating that FlaB3 likely composes the curved portion of the flagella. This could represent a unique case of a flagellin performing the role of the bacterial hook protein. FlaA-specific antibodies were used in immunoblotting to determine that FlaA is found throughout the flagellar filament. M. voltae cells were transformed with a modified flaA gene containing a hemagglutinin (HA) tag introduced into the variable region. Transformants that had replaced the wild-type copy of the flaA gene with the HA-tagged version incorporated the HA-tagged version of FlaA into flagella which appeared normal by electron microscopy.

| S-EPMC135359 | biostudies-literature

Methanococcus voltae strain:A2

Project description:Methanococcus voltae A2 genome sequencing

| PRJNA712126 | ENA

Methanococcus voltae strain:C2

Project description:Methanococcus voltae C2 genome sequencing

| PRJNA712127 | ENA

Methanococcus voltae strain:C4

Project description:Methanococcus voltae C4 genome sequencing

| PRJNA712128 | ENA