Evidence of rainbow trout prolactin interaction with its receptor through unstable homodimerisation.
This study aims to characterise Prolactin receptor (PRLR) in rainbow trout for which no information is available despite the availability of Salmonid PRL preparations. By screening a freshwater rainbow trout intestine cDNA library with a probe corresponding to the extracellular domain (ECD) of tilapia PRLR, we have cloned a 2.5 kb insert coding for the PRLR. The mature protein of 614 amino acid residues is similar to PRLR isolated in tilapia and also the long form of mammalian PRLR. Analysis of PRLR gene expression in osmoregulatory organs revealed the presence of a unique transcript, thus confirming the involvement of this hormone in the control of osmoregulation in this fish species. By using surface plasmon resonance (SPR) technology, kinetic measurement of interaction between trout PRL and its receptor ECD was studied. This approach allowed us to demonstrate the formation of a transient, unstable homodimeric complex. This unstability could explain the inability to perform binding experiments using homologous PRL. In contrast, heterologous lactogenic ligands were able to interact through a more stable complex. Whether these characteristics of PRL-receptor interaction in rainbow trout are different to what occurs in tilapia where a homologous radioreceptor assay was developed would require further studies.
Rouzic
P L
PL
Institut National de la Recherche Agronomique, Station de Recherche Commune en Ichtyophysiologie, Biodiversité et Environnement, Group on Physiology of Adaptation and Stress, campus de Beaulieu, 35042, Rennes Cedex, France.
Sandra
O
O
Grosclaude
J
J
Rentier-Delrue
F
F
Jolois
O
O
Tujague
M
M
Pakdel
F
F
Sandowski
Y
Y
Cohen
Y
Y
Gertler
A
A
Prunet
P
P
D000818:Animals; D001483:Base Sequence; D019281:Dimerization; D007700:Kinetics; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss / Q000235:genetics / Q000378:metabolism*; D011388:Prolactin / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012313:RNA / Q000378:metabolism; D011981:Receptors, Prolactin / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D020349:Surface Plasmon Resonance; D014018:Tissue Distribution; D014882:Water-Electrolyte Balance
D011981:Receptors, Prolactin; D012313:RNA; D011388:Prolactin