Binding of ligand or monoclonal antibody 4B1 induces discrete structural changes in the lactose permease of Escherichia coli
| dc.contributor.author | Frillingos, S. | en |
| dc.contributor.author | Wu, J. | en |
| dc.contributor.author | Venkatesan, P. | en |
| dc.contributor.author | Kaback, H. R. | en |
| dc.date.accessioned | 2015-11-24T19:32:56Z | |
| dc.date.available | 2015-11-24T19:32:56Z | |
| dc.identifier.issn | 0006-2960 | - |
| dc.identifier.uri | https://olympias.lib.uoi.gr/jspui/handle/123456789/23474 | |
| dc.rights | Default Licence | - |
| dc.subject | Antibodies, Monoclonal/*chemistry | en |
| dc.subject | *Binding Sites, Antibody/genetics | en |
| dc.subject | Cysteine/genetics | en |
| dc.subject | Escherichia coli/chemistry/*enzymology | en |
| dc.subject | *Escherichia coli Proteins | en |
| dc.subject | Ligands | en |
| dc.subject | Membrane Transport Proteins/*chemistry/genetics/*metabolism | en |
| dc.subject | *Monosaccharide Transport Proteins | en |
| dc.subject | Mutagenesis, Site-Directed | en |
| dc.subject | Protein Conformation | en |
| dc.subject | *Symporters | en |
| dc.title | Binding of ligand or monoclonal antibody 4B1 induces discrete structural changes in the lactose permease of Escherichia coli | en |
| heal.abstract | By using Cys-scanning mutagenesis with site-directed sulfhydryl modification in situ [Frillingos, S., & Kaback, H. R. (1996) Biochemistry 35, 3950-3956], conformational changes induced by binding of ligand or monoclonal antibody (mAb) 4B1 in the lactose permease of Escherichia coli were studied. Out of 31 single-Cys replacement mutants in helices I, V, VII, VIII, X, or XI, 4B1 binding alters the reactivity of Val238-->Cys (helix VII), Val331-->Cys (helix X), or single-Cys355 (helix XI) permease with N-ethylmaleimide (NEM) in right-side-out membrane vesicles. In addition, site-directed fluorescence spectroscopy shows that mAb 4B1 binding causes position 331 (helix X) in the permease to experience a more hydrophobic environment. In contrast, ligand binding elicits more widespread changes, as evidenced by enhancement of the NEM reactivity of Ala244-->Cys, Thr248-->Cys (helix VII), Thr265-->Cys (helix VIII), Val315-->Cys (helix X), Gln359-->Cys, or Met362-->Cys (helix XI) permease, none of which are altered by 4B1 binding. Furthermore, no effect of 4B1 is observed on the reactivity of Cys148 (helix V), Val264-->Cys, Gly268-->Cys, or Asn272-->Cys (helix VIII), positions which probably make direct contact with substrate. With respect to the N-terminal half of the permease, 4B1 binding causes a small increase in the reactivity of mutants Pro28-->Cys or Pro31-->Cys (helix I), while ligand binding causes much greater increases in reactivity. The findings indicate that 4B1 binding induces a structural change in the permease that is much less widespread than that induced by ligand binding. | en |
| heal.access | campus | - |
| heal.fullTextAvailability | TRUE | - |
| heal.identifier.primary | 10.1021/bi970233b | - |
| heal.identifier.secondary | http://www.ncbi.nlm.nih.gov/pubmed/9174357 | - |
| heal.identifier.secondary | http://pubs.acs.org/doi/pdfplus/10.1021/bi970233b | - |
| heal.journalName | Biochemistry | en |
| heal.journalType | peer-reviewed | - |
| heal.language | en | - |
| heal.publicationDate | 1997 | - |
| heal.recordProvider | Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών Υγείας. Τμήμα Ιατρικής | el |
| heal.type | journalArticle | - |
| heal.type.el | Άρθρο Περιοδικού | el |
| heal.type.en | Journal article | en |
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