Cys-scanning mutagenesis: a novel approach to structure function relationships in polytopic membrane proteins
Φόρτωση...
Ημερομηνία
Συγγραφείς
Frillingos, S.
Sahin-Toth, M.
Wu, J.
Kaback, H. R.
Τίτλος Εφημερίδας
Περιοδικό ISSN
Τίτλος τόμου
Εκδότης
Περίληψη
Τύπος
Είδος δημοσίευσης σε συνέδριο
Είδος περιοδικού
peer-reviewed
Είδος εκπαιδευτικού υλικού
Όνομα συνεδρίου
Όνομα περιοδικού
Faseb Journal
Όνομα βιβλίου
Σειρά βιβλίου
Έκδοση βιβλίου
Συμπληρωματικός/δευτερεύων τίτλος
Περιγραφή
The entire lactose permease of Escherichia coli, a polytopic membrane transport protein that catalyzes beta-galactoside/H+ symport, has been subjected to Cys-scanning mutagenesis in order to determine which residues play an obligatory role in the mechanism and to create a library of mutants with a single-Cys residue at each position of the molecule for structure/function studies. Analysis of the mutants has led to the following: 1) only six amino acid side chains play an irreplaceable role in the transport mechanism; 2) positions where the reactivity of the Cys replacement is increased upon ligand binding are identified; 3) positions where the reactivity of the Cys replacement is decreased by ligand binding are identified; 4) helix packing, helix tilt, and ligand-induced conformational changes are determined by using the library of mutants in conjunction with a battery of site-directed techniques; 5) the permease is a highly flexible molecule; and 6) a working model that explains coupling between beta-galactoside and H+ translocation. structure-function relationships in polytopic membrane proteins.
Περιγραφή
Λέξεις-κλειδιά
Alkylation, Amino Acid Sequence, Antibodies, Monoclonal/immunology, Bacterial Proteins/chemistry/*genetics, Binding Sites, Catalytic Domain, Cysteine, Escherichia coli/enzymology, *Escherichia coli Proteins, Glutamic Acid/metabolism, Ion Transport, Membrane Proteins/chemistry/*genetics, Membrane Transport Proteins/chemistry/*genetics, Models, Molecular, Molecular Sequence Data, *Monosaccharide Transport Proteins, *Mutagenesis, Site-Directed, Protein Conformation, Protein Structure, Secondary, Protons, Recombinant Fusion Proteins/chemistry/*genetics, Structure-Activity Relationship, *Symporters
Θεματική κατηγορία
Παραπομπή
Σύνδεσμος
http://www.ncbi.nlm.nih.gov/pubmed/9761772
http://www.fasebj.org/content/12/13/1281.full.pdf
http://www.fasebj.org/content/12/13/1281.full.pdf
Γλώσσα
en
Εκδίδον τμήμα/τομέας
Όνομα επιβλέποντος
Εξεταστική επιτροπή
Γενική Περιγραφή / Σχόλια
Ίδρυμα και Σχολή/Τμήμα του υποβάλλοντος
Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών Υγείας. Τμήμα Ιατρικής