Fiorito, F.

Gossert, A.

Hiller, S.

Wüthrich, K.

RERO DOC Digital Library

LETTER TO THE EDITORNMR assignment of the E. coli type 1 pilus protein FimFA. D. Gossert Æ S. Hiller Æ F. Fiorito Æ K. Wu¨thrichReceived: 27 November 2006 / Accepted: 30 November 2006 / Published online: 7 April 2007 Springer Science+Business Media B.V. 2007Type 1 pili are filamentous adhesion organelles ofuropathogenic E. coli strains, which consist of up to3,000 non-covalently connected protein subunits. Eachsubunit features an immunoglobulin-like fold, in whichone of the seven b-strands is donated by the precedingsubunit (Vetsch et al. 2005). For an NMR structuredetermination, a 179-amino acid construct of the type 1pilus subunit FimF was engineered, which contains aC-terminal extension that acts as the complementingdonor strand. Sequence-specific resonance assignmentsfor the backbone nuclei HN, 13Ca, 13Cb and 15N wereobtained using the peak lists from 4D APSY-HNCOCAand 4D APSY-COHNCA experiments (Hiller et al.2005) to support the analysis of a 3D HNCACBspectrum. This novel strategy was found to greatlyimprove the efficiency of interactive backboneresonance assignment. Side chain assignments weresupported by the automated routine ASCAN (Fioritoet al., to be published). Assignments were obtained forthe residues 2–155 and 166–179, including all HN, Ha,Hb, 13Ca, 13Cb and backbone 15N and HN atoms, 89%of the 13C¢ atoms, 97% of the side chain 13CHn moie-ties, 96% of the Asn and Gln 15NH2 groups, and 3 ofthe 6 Arg e15NH groups. (BMRB accession Nr. 15032).Financial support by NCCR Structural Biology.ReferencesVetsch et al. (2005) Protein folding handbook II:965–986Hiller S, Fiorito F, Wuthrich K, Wider G (2005) Automatedprojection spectroscopy (APSY). Proc Natl Acad Sci USA102(31):10876–10881A. D. Gossert  S. Hiller  F. Fiorito  K. Wu¨thrich (&)Institut fu¨r Molekularbiologie und Biophysik, ETH Zu¨rich,CH-8093 Zu¨rich, Switzerlande-mail: wuthrich@mol.biol.ethz.ch123J Biomol NMR (2007) 38:195DOI 10.1007/s10858-006-9123-9

NMR assignment of the E. coli type 1 pilus protein FimF

http://doc.rero.ch/record/310986/files/10858_2006_Article_9123.pdf

NMR assignment of the E. coli type 1 pilus protein FimF

Abstract

Similar works

Full text

Available Versions

RERO DOC Digital Library