Structural basis of 3-phosphoinositide recognition by pleckstrin homology domains
| dc.contributor.author | Lietzke, Susan E. | |
| dc.contributor.author | Bose, Sahana | |
| dc.contributor.author | Cronin, Thomas Charles | |
| dc.contributor.author | Klarlund, Jes K. | |
| dc.contributor.author | Chawla, Anil | |
| dc.contributor.author | Czech, Michael P. | |
| dc.contributor.author | Lambright, David G. | |
| dc.date | 2022-08-11T08:09:00.000 | |
| dc.date.accessioned | 2022-08-23T16:15:16Z | |
| dc.date.available | 2022-08-23T16:15:16Z | |
| dc.date.issued | 2000-09-13 | |
| dc.date.submitted | 2008-10-22 | |
| dc.identifier.citation | <p>Mol Cell. 2000 Aug;6(2):385-94.</p> | |
| dc.identifier.issn | 1097-2765 (Print) | |
| dc.identifier.doi | 10.1016/S1097-2765(00)00038-1 | |
| dc.identifier.pmid | 10983985 | |
| dc.identifier.uri | http://hdl.handle.net/20.500.14038/34048 | |
| dc.description.abstract | Lipid second messengers generated by phosphoinositide (PI) 3-kinases regulate diverse cellular functions through interaction with pleckstrin homology (PH) domains in modular signaling proteins. The PH domain of Grp1, a PI 3-kinase-activated exchange factor for Arf GTPases, selectively binds phosphatidylinositol 3,4,5-trisphosphate with high affinity. We have determined the structure of the Grp1 PH domain in the unliganded form and bound to inositol 1,3,4,5-tetraphosphate. A novel mode of phosphoinositide recognition involving a 20-residue insertion within the beta6/beta7 loop explains the unusually high specificity of the Grp1 PH domain and the promiscuous 3-phosphoinositide binding typical of several PH domains including that of protein kinase B. When compared to other PH domains, general determinants of 3-phosphoinositide recognition and specificity can be deduced. | |
| dc.language.iso | en_US | |
| dc.relation | <p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=10983985&dopt=Abstract">Link to article in PubMed</a></p> | |
| dc.relation.url | https://doi.org/10.1016/S1097-2765(00)00038-1 | |
| dc.subject | 1-Phosphatidylinositol 3-Kinase; Amino Acid Sequence; Amino Acid Substitution; Binding Sites; Conserved Sequence; Crystallography, X-Ray; Inositol Phosphates; Models, Molecular; Molecular Sequence Data; Mutagenesis, Site-Directed; Phosphatidylinositols; Protein Conformation; Protein Structure, Secondary; Receptors, Cytoplasmic and Nuclear; Recombinant Proteins; Sequence Alignment; Sequence Homology, Amino Acid; src Homology Domains | |
| dc.subject | Life Sciences | |
| dc.subject | Medicine and Health Sciences | |
| dc.title | Structural basis of 3-phosphoinositide recognition by pleckstrin homology domains | |
| dc.type | Journal Article | |
| dc.source.journaltitle | Molecular cell | |
| dc.source.volume | 6 | |
| dc.source.issue | 2 | |
| dc.identifier.legacycoverpage | https://escholarship.umassmed.edu/gsbs_sp/698 | |
| dc.identifier.contextkey | 654590 | |
| html.description.abstract | <p>Lipid second messengers generated by phosphoinositide (PI) 3-kinases regulate diverse cellular functions through interaction with pleckstrin homology (PH) domains in modular signaling proteins. The PH domain of Grp1, a PI 3-kinase-activated exchange factor for Arf GTPases, selectively binds phosphatidylinositol 3,4,5-trisphosphate with high affinity. We have determined the structure of the Grp1 PH domain in the unliganded form and bound to inositol 1,3,4,5-tetraphosphate. A novel mode of phosphoinositide recognition involving a 20-residue insertion within the beta6/beta7 loop explains the unusually high specificity of the Grp1 PH domain and the promiscuous 3-phosphoinositide binding typical of several PH domains including that of protein kinase B. When compared to other PH domains, general determinants of 3-phosphoinositide recognition and specificity can be deduced.</p> | |
| dc.identifier.submissionpath | gsbs_sp/698 | |
| dc.contributor.department | Program in Molecular Medicine | |
| dc.contributor.department | Graduate School of Biomedical Sciences | |
| dc.source.pages | 385-94 |
Files in this item
Name:
Publisher version
This item appears in the following Collection(s)
Related items
Showing items related by title, author, creator and subject.
-
Flagellin gene transcription in Bordetella bronchiseptica is regulated by the BvgAS virulence control systemThe products of the bvgAS locus activate expression of a majority of the known Bordetella virulence factors but also exert negative control over a class of genes called vrg genes (bvg-repressed genes). BvgAS negatively controls the production of flagella and the phenotype of motility in Bordetella bronchiseptica. In this study flaA, the flagellin gene, was cloned and characterized to facilitate studies of this negative control pathway. An internal flaA probe detected hybridizing sequences on genomic Southern blots of Bordetella pertussis, Bordetella parapertussis, and Bordetella avium, although B. pertussis and B. parapertussis are nonmotile. FlaA is similar to the FliC flagellins of Salmonella typhimurium and Escherichia coli, and flaA complemented an E. coli flagellin mutant. Insertional inactivation of the chromosomal flaA locus eliminated motility, which was restored by complementation with the wild-type locus. Analysis of flaA mRNA production by Northern (RNA) blotting and primer extension indicated that negative regulation by BvgAS occurs at the level of transcription. The transcriptional start site of flaA mapped near a consensus site for the alternative sigma factor, sigma F, encoded by fliA in E. coli and S. typhimurium. Consistent with a role for a fliA analog in B. bronchiseptica, transcriptional activation of a flaA-lacZ fusion in E. coli required fliA and a flaA-linked locus designated frl.frl also efficiently complemented mutations in the flagellar master regulatory locus, flhDC, of E. coli. Our analysis of the motility phenotype of B. bronchiseptica suggests that the Bordetella virulence control system mediates transcriptional control of flaA through a regulatory hierarchy that includes the frl locus and an alternative sigma factor.
-
The unique catalytic subunit of sperm cAMP-dependent protein kinase is the product of an alternative Calpha mRNA expressed specifically in spermatogenic cellscAMP-dependent protein kinase has a central role in the control of mammalian sperm capacitation and motility. Previous protein biochemical studies indicated that the only cAMP-dependent protein kinase catalytic subunit (C) in ovine sperm is an unusual isoform, termed C(s), whose amino terminus differs from those of published C isoforms of other species. Isolation and sequencing of cDNA clones encoding ovine C(s) and Calpha1 (the predominant somatic isoform) now reveal that C(s) is the product of an alternative transcript of the Calpha gene. C(s) cDNA clones from murine and human testes also were isolated and sequenced, indicating that C(s) is of ancient origin and widespread in mammals. In the mouse, C(s) transcripts were detected only in testis and not in any other tissue examined, including ciliated tissues and ovaries. Finally, immunohistochemistry of the testis shows that C(s) first appears in pachytene spermatocytes. This is the first demonstration of a cell type-specific expression for any C isoform. The conservation of C(s) throughout mammalian evolution suggests that the unique structure of C(s) is important in the subunit's localization or function within the sperm.
-
Moesin, ezrin, and p205 are actin-binding proteins associated with neutrophil plasma membranesActin-binding proteins in bovine neutrophil plasma membranes were identified using blot overlays with 125I-labeled F-actin. Along with surface-biotinylated proteins, membranes were enriched in major actin-binding polypeptides of 78, 81, and 205 kDa. Binding was specific for F-actin because G-actin did not bind. Further, unlabeled F-actin blocked the binding of 125I-labeled F-actin whereas other acidic biopolymers were relatively ineffective. Binding also was specifically inhibited by myosin subfragment 1, but not by CapZ or plasma gelsolin, suggesting that the membrane proteins, like myosin, bind along the sides of the actin filaments. The 78- and 81-kDa polypeptides were identified as moesin and ezrin, respectively, by co-migration on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoprecipitation with antibodies specific for moesin and ezrin. Although not present in detectable amounts in bovine neutrophils, radixin (a third and closely related member of this gene family) also bound 125I-labeled F-actin on blot overlays. Experiments with full-length and truncated bacterial fusion proteins localized the actin-binding site in moesin to the extreme carboxy terminus, a highly conserved sequence. Immunofluorescence micrographs of permeabilized cells and cell "footprints" showed moesin co-localization with actin at the cytoplasmic surface of the plasma membrane, consistent with a role as a membrane-actin-linking protein.
