ArticleJournal of microbiological methods2008
The utility of affinity-tags for detection of a streptococcal protein from a variety of streptococcal species.
Article in Journal of microbiological methods, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
21 citing papers in PubMed, 25 citations in OpenAlex.
- In Situ Biofilm Affinity-Based Protein Profiling Identifies the Streptococcal Hydrolase GbpB as the Target of a Carolacton-Inspired Chemical Probe.Journal of the American Chemical Society · 2024Article
- Intraspecies interactions ofmSphere · 2024Article
- Illuminating the oral microbiome and its host interactions: tools and approaches for molecular microbiology studies.FEMS microbiology reviews · 2023Review
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- Caries-Associated Biosynthetic Gene Clusters inJournal of dental research · 2020Article
- Dietary Nitrite Drives Disease Outcomes in Oral Polymicrobial Infections.Journal of dental research · 2019Article
- Glycosyltransferase-Mediated Biofilm Matrix Dynamics and Virulence of Streptococcus mutans.Applied and environmental microbiology · 2019Article
- Antigen I/II mediates interactions between Streptococcus mutans and Candida albicans.Molecular oral microbiology · 2018Article
- Engineering and Dissecting the Glycosylation Pathway of a Streptococcal Serine-rich Repeat Adhesin.The Journal of biological chemistry · 2016Article
- Streptococcus mutans copper chaperone, CopZ, is critical for biofilm formation and competitiveness.Molecular oral microbiology · 2016Article
- New Helical Binding Domain Mediates a Glycosyltransferase Activity of a Bifunctional Protein.The Journal of biological chemistry · 2016Article
- A distinct sortase SrtB anchors and processes a streptococcal adhesin AbpA with a novel structural property.Scientific reports · 2016Article
- Cyclic di-AMP mediates biofilm formation.Molecular microbiology · 2016Article
- A conserved domain is crucial for acceptor substrate binding in a family of glucosyltransferases.Journal of bacteriology · 2015Article
- YlxM is a newly identified accessory protein that influences the function of signal recognition particle pathway components in Streptococcus mutans.Journal of bacteriology · 2014Article
- Gap2 promotes the formation of a stable protein complex required for mature Fap1 biogenesis.Journal of bacteriology · 2013Article
- Gap1 functions as a molecular chaperone to stabilize its interactive partner Gap3 during biogenesis of serine-rich repeat bacterial adhesin.Molecular microbiology · 2012Article
- A molecular chaperone mediates a two-protein enzyme complex and glycosylation of serine-rich streptococcal adhesins.The Journal of biological chemistry · 2011Article
- New cell surface protein involved in biofilm formation by Streptococcus parasanguinis.Infection and immunity · 2011Article
- Purification and characterization of an active N-acetylglucosaminyltransferase enzyme complex from Streptococci.Applied and environmental microbiology · 2010Article
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
Abstract
There is no systematic examination of affinity tag utility in Gram-positive bacteria, which limits the investigation of protein function in this important group of bacteria as specific antibodies for many of native proteins are generally not available. In this study, we utilized an E. coli-streptococcal shuttle vector pVT1666 and constructed two sets of expression plasmids pVPT-CTag and pVPT-NTag, with each set containing five affinity tags (GST, GFP, HSV, T7 and Nano) that can be fused to either the C- or N-terminus of a target protein. A putative glycosyltransferase (Gtf2) essential for Fap1 glycosylation was used to demonstrate the utility of the cassettes in detection of Gtf2 fusion proteins, and the biological relevance of the proteins in our working strain Streptococcus parasanguinis. GFP and T7 tags were readily expressed in S. parasanguinis as either an N- or C-terminal fusion to Gtf2. Only the C- terminal fusion of GST and HSV were able to be identified in S. parasanguinis. The Nano tag was not detected in either E. coli or S. parasanguinis. Genetic complementation experiments indicated that all the tagged Gtf2 fusion proteins could restore the Gtf2 function in the null mutant except for the Nano-tagged Gtf2 at its N-terminal fusion. Using a T7-tagged Gtf2 fusion construct, we demonstrated that the fusion cassette is also useful in detection of the fusion tag expression in other streptococci including S. mutans, S. pneumoniae and S. sanguinis. Therefore, the expression cassettes we constructed will be a useful tool not only to investigate protein-protein interactions in Fap1 biogenesis in S. parasanguinis, but also to study protein functions in other gram-positive bacteria in which pVT1666 replicates.
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