Mapping spatial approximations between the amino terminus of secretin and each of the extracellular loops of its receptor using cysteine trapping

被引：31

作者：

Dong, Maoqing ^{[1
]}

Xu, Xiequn ^{[1
]}

Ball, Alicja M. ^{[1
]}

Makhoul, Joshua A. ^{[1
]}

Lam, Polo C. -H. ^{[2
]}

Pinon, Delia I. ^{[1
]}

Orry, Andrew ^{[2
]}

Sexton, Patrick M. ^{[3
,4
]}

Abagyan, Ruben ^{[2
,5
]}

Miller, Laurence J. ^{[1
]}

机构：

[1] Mayo Clin, Dept Mol Pharmacol & Expt Therapeut, Scottsdale, AZ 85259 USA

[2] Molsoft LLC, La Jolla, CA USA

[3] Monash Univ, Monash Inst Pharmaceut Sci, Parkville, Vic, Australia

[4] Monash Univ, Dept Pharmacol, Parkville, Vic, Australia

[5] Univ Calif San Diego, Skaggs Sch Pharm & Pharmaceut Sci, La Jolla, CA 92093 USA

来源：

FASEB JOURNAL | 2012年 / 26卷 / 12期

基金：

美国国家卫生研究院; 英国医学研究理事会;

关键词：

class B GPCRs; molecular modeling; ligand binding; PHOTOAFFINITY CROSS-LINKING; PROTEIN-COUPLED-RECEPTOR; 2ND TRANSMEMBRANE HELIX; PEPTIDE RECEPTOR; ANTAGONIST PEPTIDES; MUTATIONAL ANALYSIS; AGONIST BINDING; MOLECULAR-BASIS; LIGAND-BINDING; BASIC RESIDUES;

D O I：

10.1096/fj.12-212399

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

While it is evident that the carboxylterminal region of natural peptide ligands bind to the amino-terminal domain of class B GPCRs, how their biologically critical amino-terminal regions dock to the receptor is unclear. We utilize cysteine trapping to systematically explore spatial approximations among residues in the first five positions of secretin and in every position within the receptor extracellular loops (ECLs). Only Cys(2) and Cys(5) secretin analogues exhibited full activity and retained moderate binding affinity (IC50: 92+/-4 and 83+/-1 nM, respectively). When these peptides probed 61 human secretin receptor cysteine-replacement mutants, a broad network of receptor residues could form disulfide bonds consistent with a dynamic ligand-receptor interface. Two distinct patterns of disulfide bond formation were observed: Cys2 predominantly labeled residues in the amino terminus of ECL2 and ECL3 (relative labeling intensity: Ser(340), 94+/-7%; Pro(341), 84+/-9%; Phe(258), 73+/-5%; Trp(274) 62+/-8%), and Cys(5) labeled those in the carboxyl terminus of ECL2 and ECL3 (Gln(348), 100%; Ile(347), 73+/-12%; Glu(342), 59+/-10%; Phe(351), 58+/-11%). These constraints were utilized in molecular modeling, providing improved understanding of the structure of the transmembrane bundle and interconnecting loops, the orientation between receptor domains, and the molecular basis of ligand docking. Key spatial approximations between peptide and receptor predicted by this model (H-1-W-274, D-3-N-268, G(4)-F-258) were supported by mutagenesis and residue-residue complementation studies.-Dong, M., Xu, X., Ball, A. M., Makhoul, J. A., Lam, P. C.-H., Pinon, D. I., Orry, A., Sexton, P. M., Abagyan, R., Miller, L. J. Mapping spatial approximations between the amino terminus of secretin and each of the extracellular loops of its receptor using cysteine trapping. FASEB J. 26, 5092-5105 (2012). www.fasebj.org

引用

页码：5092 / 5105

页数：14

共 13 条

[1] Mapping spatial approximations between residues contributing to the helix N-capping motif of secretin and distinct residues within each of the extracellular loops of its receptor using cysteine trapping.
Dong, M.
Pinon, D. I.
Miller, L. J.
MOLECULAR BIOLOGY OF THE CELL, 2012, 23
[2] Use of Cysteine Trapping to Map Spatial Approximations between Residues Contributing to the Helix N-capping Motif of Secretin and Distinct Residues within Each of the Extracellular Loops of Its Receptor
Dong, Maoqing
Lam, Polo C. -H.
Orry, Andrew
Sexton, Patrick M.
Christopoulos, Arthur
Abagyan, Ruben
Miller, Laurence J.
JOURNAL OF BIOLOGICAL CHEMISTRY, 2016, 291 (10) : 5172 - 5184
[3] Molecular approximations between residues 21 and 23 of secretin and its receptor: Development of a model for peptide docking with the amino terminus of the secretin receptor
Dong, Maoqing
Lam, Polo C. -H.
Gao, Fan
Hosohata, Keiko
Pinon, Delia I.
Sexton, Patrick M.
Abagyan, Ruben
Miller, Laurence J.
MOLECULAR PHARMACOLOGY, 2007, 72 (02) : 280 - 290
[4] Mapping Interactions Between the Amino-Terminal Region of Secretin and its Receptor using Disulfide-Trapping
Dong, Maoqing
Xu, Xiequn
Ball, Alicja
Makhoul, Joshua A.
Lam, Polo P. C.
Pinon, Delia I.
Sexton, Patrick M.
Abagyan, Ruben
Miller, Laurence J.
BIOPHYSICAL JOURNAL, 2012, 102 (03) : 515A - 515A
[5] Spatial approximation between a photolabile residue in position 13 of secretin and the amino terminus of the secretin receptor
Zang, MW
Dong, MQ
Pinon, DI
Ding, XQ
Hadac, EM
Li, ZJ
Lybrand, TP
Miller, LJ
MOLECULAR PHARMACOLOGY, 2003, 63 (05) : 993 - 1001
[6] Demonstration of approximation between the amino terminus of secretin and transmembrane segment six of its receptor
Dong, M
Pinon, DI
Miller, LJ
MOLECULAR BIOLOGY OF THE CELL, 2002, 13 : 80A - 80A
[7] Spatial approximation between the amino terminus of a peptide agonist and the top of the sixth transmembrane segment of the secretin receptor
Dong, MQ
Li, ZJ
Pinon, DI
Lybrand, TP
Miller, LJ
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (04) : 2894 - 2903
[8] Spatial approximation between a photolabile residue in position 13 of secretin and the amino terminus of the secretin receptor (vol 63, pg 993, 2003)
Zang, M
Dong, M
Pinon, DI
Ding, XQ
Hadac, EM
Li, Z
Lybrand, TP
Miller, LJ
MOLECULAR PHARMACOLOGY, 2003, 64 (01) : 193 - 193
[9] Differential spatial approximations between secretin and its receptor in active and inactive conformations demonstrated by photoaffinity labeling
Hosohata, K
Dong, M
Pinon, DI
Miller, LJ
REGULATORY PEPTIDES, 2005, 130 (03) : 165 - 165
[10] Spatial approximation between two residues in the mid-region of secretin and the amino terminus of its receptor - Incorporation of seven sets of such constraints into a three-dimensional model of the agonist-bound secretin receptor
Dong, MQ
Li, ZJ
Zang, MQ
Pinon, DI
Lybrand, TP
Miller, LJ
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (48) : 48300 - 48312

← 1 2 →