TY - JOUR
T1 - Evidence of rainbow trout prolactin interaction with its receptor through unstable homodimerisation
AU - Rouzic, Philippe Le
AU - Sandra, Olivier
AU - Grosclaude, Jeanne
AU - Rentier-Delrue, Françoise
AU - Jolois, Olivier
AU - Tujague, Michel
AU - Pakdel, Farzad
AU - Sandowski, Yael
AU - Cohen, Yael
AU - Gertler, Arieh
AU - Prunet, Patrick
PY - 2001/2/14
Y1 - 2001/2/14
N2 - 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.
AB - 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.
KW - Gene expression
KW - Osmoregulation
KW - Prolactin receptor
KW - Receptor extracellular domain
KW - Surface plasmon resonance (SPR)
KW - cDNA cloning
UR - http://www.scopus.com/inward/record.url?scp=0035857439&partnerID=8YFLogxK
U2 - 10.1016/S0303-7207(00)00377-4
DO - 10.1016/S0303-7207(00)00377-4
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C2 - 11165044
AN - SCOPUS:0035857439
SN - 0303-7207
VL - 172
SP - 105
EP - 113
JO - Molecular and Cellular Endocrinology
JF - Molecular and Cellular Endocrinology
IS - 1-2
ER -