PDBsum entry: 1nzs

Signaling protein

PDB id: 1nzs

Contents

Protein chain

19 a.a.

PDB id:

1nzs

Name:

Signaling protein

Title:

Nmr structures of phosphorylated carboxy terminus of bovine rhodopsin in arrestin-bound state

Structure:

19-mer peptide fragment of rhodopsin. Chain: a. Fragment: c-terminal domain, residues 330-348. Engineered: yes. Mutation: yes. Other_details: all ser and thr phosphorylated

Source:

Synthetic: yes. Other_details: this sequence occurs naturally in bos taurus. In a synthetic peptide, all serines and threonines are phosphorylated.

NMR struc:

16 models

Authors:

O.G.Kisselev,J.H.Mcdowell,P.A.Hargrave

Key ref:

O.G.Kisselev et al. (2004). The arrestin-bound conformation and dynamics of the phosphorylated carboxy-terminal region of rhodopsin. FEBS Lett, 564, 307-311. PubMed id: 15111114 DOI: 10.1016/S0014-5793(04)00226-1

Date:

19-Feb-03 Release date: 02-Mar-04

Protein chain

P02699  (OPSD_BOVIN) -  Rhodopsin

Seq: 348 a.a.

Struc: 19 a.a.*

* PDB and UniProt seqs differ at 4 residue positions (black crosses)

DOI no: 10.1016/S0014-5793(04)00226-1

FEBS Lett 564:307-311 (2004)

PubMed id: 15111114

The arrestin-bound conformation and dynamics of the phosphorylated carboxy-terminal region of rhodopsin.

O.G.Kisselev, J.H.McDowell, P.A.Hargrave.

ABSTRACT

Visual arrestin binds to the phosphorylated carboxy-terminal region of rhodopsin to block interactions with transducin and terminate signaling in the rod photoreceptor cells. A synthetic seven-phospho-peptide from the C-terminal region of rhodopsin, Rh(330-348), has been shown to bind arrestin and mimic inhibition of signal transduction. In this study, we examine conformational changes in this synthetic peptide upon binding to arrestin by high-resolution proton nuclear magnetic resonance (NMR). We show that the peptide is completely disordered in solution, but becomes structured upon binding to arrestin. A control, unphosphorylated peptide that fails to bind to arrestin remains highly disordered. Specific NMR distance constraints are used to model the arrestin-bound conformation. The models suggest that the phosphorylated carboxy-terminal region of rhodopsin, Rh(330-348), undergoes significant conformational changes and becomes structured upon binding to arrestin.

Selected figure(s)

Figure 1.
Fig. 1. Sections of ^1H-TOCSY (blue) and ^1H-NOESY spectra of 7PP in the absence (red) and in the presence (black) of bovine arrestin. The amino acid sequence of 7PP is shown. Serine and threonine residues are phosphorylated.

Figure 2.
Fig. 2. NMR structures of the phosphorylated C-terminal domain of the bovine rhodopsin in the arrestin-bound state. a: Summary of the experimental distance constraints. Black bar, intraresidue; gray, sequential; white, long-range NOEs. b: Root mean square deviation, RMSD, between individual NMR structures in the final ensemble. Black bar, main chain atoms; white bar, side chain atoms. c: Structural statistics determined by PROCHECK-NMR. d: Lowest energy NMR structure of 7PP, 7-phospho-Rh(330–348), in yellow as part of an X-ray structure of rhodopsin [19] shown as a ribbon model. The N- and C-termini of rhodopsin are labeled. Inset: Final ensemble of the 15 NMR structures superimposed using main chain atoms. There was no NMR evidence of arrestin-induced conformational changes for a control unphosphorylated version of 7PP studied under the same experimental conditions.

The above figures are reprinted by permission from the Federation of European Biochemical Societies: FEBS Lett (2004, 564, 307-311) copyright 2004.