Article (Scientific journals)
Identification Of Key Residues For Interaction Of Vasoactive Intestinal Peptide With Human Vpac(1) And Vpac(2) Receptors And Development Of A Highly Selective Vpac(1) Receptor Agonist - Alanine Scanning And Molecular Modeling Of The Peptide
Nicole, P.; Lins, Laurence; Rouyer-Fessard, C. et al.
2000In Journal of Biological Chemistry, 275 (31), p. 24003-24012
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Abstract :
[en] The widespread neuropeptide vasoactive intestinal peptide (VIP) has two receptors VPAC(1) and VPAC(2). Solid-phase syntheses of VIP analogs in which each amino acid has been changed to alanine (Ala scan) or glycine was achieved and each analog was tested for: (i) three-dimensional structure by ab initio molecular modeling; (ii) ability to inhibit (125)I-VIP binding (K(i)) and to stimulate adenylyl cyclase activity (EC(50)) in membranes from cell clones stably expressing human recombinant VPAC(1) or VPAC(2) receptor. The data show that substituting residues at 14 positions out of 28 in VIP resulted in a >10-fold increase of K(i) or EC(50) at the VPAC(1) receptor. Modeling of the three-dimensional structure of native VIP (central alpha-helice from Val(5) to Asn(24) with random coiled N and C terminus) and analogs shows that substitutions of His(1), Val(5), Arg(14), Lys(15), Lys(21), Leu(23), and Ile(26) decreased biological activity without altering the predicted structure, supporting that those residues directly interact with VPAC(1) receptor. The interaction of the analogs with human VPAC(2) receptor is similar to that observed with VPAC(1) receptor, with three remarkable exceptions: substitution of Thr(11) and Asn(28) by alanine increased K(i) for binding to VPAC(2) receptor; substitution of Tyr(22) by alanine increased EC(50) for stimulating adenylyl cyclase activity through interaction with the VPAC(2) receptor. By combining 3 mutations at positions 11, 22, and 28, we developed the [Ala(11,22,28)]VIP analog which constitutes the first highly selective (>1,000-fold) human VPAC(1) receptor agonist derived from VIP ever described.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Nicole, P.
Lins, Laurence  ;  Université de Liège - ULiège > Gembloux Agro-Bio Tech
Rouyer-Fessard, C.
Drouot, C.
Fulcrand, P.
Thomas, Annick ;  Université de Liège - ULiège > Chimie et bio-industries > Centre de Bio. Fond. - Section de Biologie moléc. et numér.
Couvineau, A.
Martinez, J.
Brasseur, Robert ;  Université de Liège - ULiège > Gembloux Agro-Bio Tech
Laburthe, M.
Language :
English
Title :
Identification Of Key Residues For Interaction Of Vasoactive Intestinal Peptide With Human Vpac(1) And Vpac(2) Receptors And Development Of A Highly Selective Vpac(1) Receptor Agonist - Alanine Scanning And Molecular Modeling Of The Peptide
Publication date :
2000
Journal title :
Journal of Biological Chemistry
ISSN :
0021-9258
eISSN :
1083-351X
Publisher :
American Society for Biochemistry and Molecular Biology, United States - Maryland
Volume :
275
Issue :
31
Pages :
24003-24012
24003-12
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 25 June 2010

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