PDBsum entry 4xee

Signaling protein, hydrolase

PDB id: 4xee

Contents

Protein chain

464 a.a.

Ligands

ARG-ARG-PRO-TYR-ILE-LEU

1PE ×2

EPE

FLC

PEG

Waters ×10

PDB id:

4xee

Name:

Signaling protein, hydrolase

Title:

Structure of active-like neurotensin receptor

Structure:

Neurotensin receptor type 1, endolysin chimera. Chain: a. Fragment: unp residues 43-396 (p20789), residues 2-161 (p00720). Synonym: ntr1, high-affinity levocabastine-insensitive neurotensin receptor, ntrh,lysis protein, lysozyme, muramidase. Engineered: yes. Mutation: yes. Neurotensin/neuromedin n. Chain: b.

Source:

Rattus norvegicus, enterobacteria phage t4. Rat. Organism_taxid: 10116, 10665. Gene: ntsr1, ntsr. Expressed in: trichoplusia ni. Expression_system_taxid: 7111. Synthetic: yes. Rattus norvegicus. Organism_taxid: 10116

Resolution:

2.90Å R-factor: 0.234 R-free: 0.281

Authors:

B.E.Krumm,J.F.White,P.Shah,R.Grisshammer

Key ref:

B.E.Krumm et al. (2015). Structural prerequisites for G-protein activation by the neurotensin receptor. Nat Commun, 6, 7895. PubMed id: 26205105 DOI: 10.1038/ncomms8895

Date:

23-Dec-14 Release date: 29-Jul-15

Protein chain

P00720  (ENLYS_BPT4) -  Endolysin from Enterobacteria phage T4

Seq: 164 a.a.

Struc: 464 a.a.*

Protein chain

P20789  (NTR1_RAT) -  Neurotensin receptor type 1 from Rattus norvegicus

Seq: 424 a.a.

Struc: 464 a.a.*

* PDB and UniProt seqs differ at 53 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.3.2.1.17 - lysozyme.
• Reaction: Hydrolysis of the 1,4-beta-linkages between N-acetyl-D-glucosamine and N-acetylmuramic acid in peptidoglycan heteropolymers of the prokaryotes cell walls.

DOI no: 10.1038/ncomms8895

Nat Commun 6:7895 (2015)

PubMed id: 26205105

Structural prerequisites for G-protein activation by the neurotensin receptor.

B.E.Krumm, J.F.White, P.Shah, R.Grisshammer.

ABSTRACT

We previously determined the structure of neurotensin receptor NTSR1 in an active-like conformation with six thermostabilizing mutations bound to the peptide agonist neurotensin. This receptor was unable to activate G proteins, indicating that the mutations restricted NTSR1 to relate agonist binding to G-protein activation. Here we analyse the effect of three of those mutations (E166A(3.49), L310A(6.37), F358A(7.42)) and present two structures of NTSR1 able to catalyse nucleotide exchange at Gα. The presence of F358(7.42) causes the conserved W321(6.48) to adopt a side chain orientation parallel to the lipid bilayer sealing the collapsed Na(+) ion pocket and linking the agonist with residues in the lower receptor part implicated in GPCR activation. In the intracellular receptor half, the bulkier L310(6.37) side chain dictates the position of R167(3.50) of the highly conserved D/ERY motif. These residues, together with the presence of E166(3.49) provide determinants for G-protein activation by NTSR1.