PDBsum entry 4zwj

Signaling protein

PDB id: 4zwj

Contents

Protein chains

833 a.a.

673 a.a.

789 a.a.

PDB id:

4zwj

Name:

Signaling protein

Title:

Crystal structure of rhodopsin bound to arrestin by femtosecond x-ray laser

Structure:

Chimera protein of human rhodopsin, mouse s-arrestin, and t4 endolysin. Chain: a, b, c, d. Synonym: lysis protein,lysozyme,muramidase,opsin-2,48 kda protein, retinal s-antigen,s-ag,rod photoreceptor arrestin. Engineered: yes. Mutation: yes

Source:

Enterobacteria phage t4, homo sapiens, mus musculus. Human, mouse. Organism_taxid: 10665, 9606, 10090. Cell_line: hek293s. Gene: rho, opn2, sag. Expressed in: homo sapiens. Expression_system_taxid: 9606. Expression_system_cell_line: hek293s

Resolution:

3.30Å R-factor: 0.255 R-free: 0.293

Authors:

Y.Kang,X.E.Zhou,X.Gao,Y.He,W.Liu,A.Ishchenko,A.Barty,T.A.White, O.Yefanov,G.W.Han,Q.Xu,P.W.De Waal,J.Ke,M.H.E.Tan,C.Zhang,A.Moeller, G.M.West,B.Pascal,N.V.Eps,L.N.Caro,S.A.Vishnivetskiy,R.J.Lee, K.M.Suino-Powell,X.Gu,K.Pal,J.Ma,X.Zhi,S.Boutet,G.J.Williams, M.Messerschmidt,C.Gati,N.A.Zatsepin,D.Wang,D.James,S.Basu,S.Roy- Chowdhury,C.Conrad,J.Coe,H.Liu,S.Lisova,C.Kupitz,I.Grotjohann, R.Fromme,Y.Jiang,M.Tan,H.Yang,J.Li,M.Wang,Z.Zheng,D.Li,N.Howe, Y.Zhao,J.Standfuss,K.Diederichs,Y.Dong,C.S.Potter,B.Carragher, M.Caffrey,H.Jiang,H.N.Chapman,J.C.H.Spence,P.Fromme,U.Weierstall,

Key ref:

Y.Kang et al. (2015). Crystal structure of rhodopsin bound to arrestin by femtosecond X-ray laser. Nature, 523, 561-567. PubMed id: 26200343 DOI: 10.1038/nature14656

Date:

19-May-15 Release date: 29-Jul-15

Protein chains

P00720  (ENLYS_BPT4) -  Endolysin from Enterobacteria phage T4

Seq: 164 a.a.

Struc: 833 a.a.*

Protein chains

P08100  (OPSD_HUMAN) -  Rhodopsin from Homo sapiens

Seq: 348 a.a.

Struc: 833 a.a.*

Protein chains

P20443  (ARRS_MOUSE) -  S-arrestin from Mus musculus

Seq: 403 a.a.

Struc: 833 a.a.*

Protein chain

P00720  (ENLYS_BPT4) -  Endolysin from Enterobacteria phage T4

Seq: 164 a.a.

Struc: 673 a.a.*

Protein chain

P08100  (OPSD_HUMAN) -  Rhodopsin from Homo sapiens

Seq: 348 a.a.

Struc: 673 a.a.*

Protein chain

P20443  (ARRS_MOUSE) -  S-arrestin from Mus musculus

Seq: 403 a.a.

Struc: 673 a.a.*

Protein chain

P00720  (ENLYS_BPT4) -  Endolysin from Enterobacteria phage T4

Seq: 164 a.a.

Struc: 789 a.a.*

Protein chain

P08100  (OPSD_HUMAN) -  Rhodopsin from Homo sapiens

Seq: 348 a.a.

Struc: 789 a.a.*

Protein chain

P20443  (ARRS_MOUSE) -  S-arrestin from Mus musculus

Seq: 403 a.a.

Struc: 789 a.a.*

* PDB and UniProt seqs differ at 255 residue positions (black crosses)

Enzyme reactions

• Enzyme class: Chains A, B, C, D: 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/nature14656

Nature 523:561-567 (2015)

PubMed id: 26200343

Crystal structure of rhodopsin bound to arrestin by femtosecond X-ray laser.

Y.Kang, X.E.Zhou, X.Gao, Y.He, W.Liu, A.Ishchenko, A.Barty, T.A.White, O.Yefanov, G.W.Han, Q.Xu, P.W.de Waal, J.Ke, M.H.Tan, C.Zhang, A.Moeller, G.M.West, B.D.Pascal, N.Van Eps, L.N.Caro, S.A.Vishnivetskiy, R.J.Lee, K.M.Suino-Powell, X.Gu, K.Pal, J.Ma, X.Zhi, S.Boutet, G.J.Williams, M.Messerschmidt, C.Gati, N.A.Zatsepin, D.Wang, D.James, S.Basu, S.Roy-Chowdhury, C.E.Conrad, J.Coe, H.Liu, S.Lisova, C.Kupitz, I.Grotjohann, R.Fromme, Y.Jiang, M.Tan, H.Yang, J.Li, M.Wang, Z.Zheng, D.Li, N.Howe, Y.Zhao, J.Standfuss, K.Diederichs, Y.Dong, C.S.Potter, B.Carragher, M.Caffrey, H.Jiang, H.N.Chapman, J.C.Spence, P.Fromme, U.Weierstall, O.P.Ernst, V.Katritch, V.V.Gurevich, P.R.Griffin, W.L.Hubbell, R.C.Stevens, V.Cherezov, K.Melcher, H.E.Xu.

ABSTRACT

G-protein-coupled receptors (GPCRs) signal primarily through G proteins or arrestins. Arrestin binding to GPCRs blocks G protein interaction and redirects signalling to numerous G-protein-independent pathways. Here we report the crystal structure of a constitutively active form of human rhodopsin bound to a pre-activated form of the mouse visual arrestin, determined by serial femtosecond X-ray laser crystallography. Together with extensive biochemical and mutagenesis data, the structure reveals an overall architecture of the rhodopsin-arrestin assembly in which rhodopsin uses distinct structural elements, including transmembrane helix 7 and helix 8, to recruit arrestin. Correspondingly, arrestin adopts the pre-activated conformation, with a ∼20° rotation between the amino and carboxy domains, which opens up a cleft in arrestin to accommodate a short helix formed by the second intracellular loop of rhodopsin. This structure provides a basis for understanding GPCR-mediated arrestin-biased signalling and demonstrates the power of X-ray lasers for advancing the frontiers of structural biology.