Prolactin regulation of the calmodulin-dependent protein kinase III elongation factor-2 system in the rat corpus luteum.
A M(r) 100,000 phosphoprotein in the corpus luteum was identified as elongation factor 2 (EF-2). Since prolactin (PRL) is necessary for optimal luteal development and protein synthesis, we determined whether this hormone affects the content and/or phosphorylation of EF-2 in the corpus luteum. PRL treatment enhanced the Ca2+/calmodulin (CaM)-dependent phosphorylation of endogenous EF-2 in luteal cytoplasmic extracts. Immunoblot analysis revealed that PRL had no effect on EF-2 levels, but examination of luteal EF-2 by two-dimensional isoelectric focusing/SDS-polyacrylamide gel electrophoresis showed that PRL increased the relative amount of the most basic dephosphorylated forms of EF-2. This suggests that PRL induces net dephosphorylation of the protein in vivo. Since EF-2 phosphorylation is regulated by both Ca2+/CaM-dependent kinase III (CaM kinase III) and protein phosphatase 2A, we examined the effect of PRL on both enzymes. Paradoxically, PRL enhanced the in vitro activity of CaM kinase III, possibly reflecting increased kinase levels, but had no effect on phosphatase activity. These results suggest that PRL maintains luteal EF-2 in a relatively dephosphorylated state in vivo by limiting the availability of Ca2+ and/or CaM to CaM kinase III. These data provide strong evidence for a role of the EF-2/CaM kinase III system in PRL action in the corpus luteum.
Albarracin
C T
CT
Department of Physiology and Biophysics, University of Illinois, Chicago 60612.
Palfrey
H C
HC
Duan
W R
WR
Rao
M C
MC
Gibori
G
G
D000818:Animals; D000906:Antibodies; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D003338:Corpus Luteum / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D054736:Elongation Factor 2 Kinase; D005260:Female; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors / Q000276:immunology / Q000378:metabolism*; D010766:Phosphorylation; D011247:Pregnancy; D011388:Prolactin / Q000494:pharmacology*; D011506:Proteins / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D000906:Antibodies; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors; D011506:Proteins; D011388:Prolactin; C517691:Eef2k protein, rat; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D054736:Elongation Factor 2 Kinase