Article (Scientific journals)
Characterization of lactogen receptor-binding site 1 of human prolactin
Kinet, Sandrina; Goffin, Vincent; Mainfroid, Véronique et al.
1996In Journal of Biological Chemistry, 271 (24), p. 14353-60
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Keywords :
Animals; Base Sequence; Binding Sites; Binding, Competitive; Cell Division/drug effects; Circular Dichroism; Computer Simulation; Humans; Kinetics; Lymphoma; Models, Structural; Molecular Sequence Data; Molecular Weight; Mutagenesis, Site-Directed; Oligodeoxyribonucleotides; Point Mutation; Prolactin/*chemistry/*metabolism/pharmacology; *Protein Structure, Secondary; Receptors, Peptide/*metabolism; Receptors, Prolactin/*metabolism; Recombinant Proteins/chemistry/metabolism/pharmacology; Software; Tumor Cells, Cultured
Abstract :
[en] Prolactin (PRL) binds to two molecules of PRL receptor (PRLR) through two regions referred to as binding sites 1 and 2. Although binding site 1 has been generally assigned to the pocket delimited by helix 1, helix 4, and the second half of loop 1, the residues involved in receptor binding have not yet all been precisely identified. In an earlier alanine-scanning mutational study, we identified three major binding determinants in loop 1 of human PRL (hPRL) (Goffin, V., Norman, M. & Martial, J. A.(1992) Mol. Endocrinol. 6, 1381-1392). Here we focus on the two other regions that form binding site 1, namely helices 1 and 4. Putative binding residues, selected on the basis of a three-dimensional model of hPRL constructed in this laboratory, were mutated to alanine, and recombinant hPRL mutants produced in Escherichia coli were tested for their ability to bind to the PRLR and to stimulate Nb2 cell proliferation. We thus identified nine single mutations (three in helix 1 and six in helix 4) whose effect was to reduce both binding and mitogenic activity by more than half as compared with wild-type hPRL, indicating the functional involvement of the corresponding residues. Adding these to the three binding determinants identified in loop 1, we now propose a complete picture of PRLR-binding site 1 of hPRL. As we earlier hypothesized, the binding site 1 determinants of hPRL differ from those of human growth hormone, a hPRL homolog.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Kinet, Sandrina;  Université de Liège - ULiège
Goffin, Vincent;  Université de Liège - ULiège
Mainfroid, Véronique;  Université de Liège - ULiège
Martial, Joseph ;  Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire
Language :
English
Title :
Characterization of lactogen receptor-binding site 1 of human prolactin
Publication date :
1996
Journal title :
Journal of Biological Chemistry
ISSN :
0021-9258
eISSN :
1083-351X
Publisher :
American Society for Biochemistry and Molecular Biology, Baltimore, United States - Maryland
Volume :
271
Issue :
24
Pages :
14353-60
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 04 December 2009

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