Uterine milk protein, a novel activin-binding protein, is present in ovine allantoic fluid.
Activins are pluripotent growth factors that have recently been shown to be present in placental and fetal membrane preparations. Our previous studies have identified and purified activin A from ovine amniotic and allantoic fluids. In this study, ligand blots of side fractions from the isolation of activin A from allantoic fluid suggested the presence of activin-binding proteins other than follistatin. Further purification of one of these fractions involved two sequential reverse phase HPLC steps and a Superose 12HR fractionation. SDS-PAGE revealed a single protein band of 55 kDa, which was identified by NH2-terminal sequencing as ovine uterine milk protein (UTMP), a member of the serine protease inhibitor (serpin) superfamily of proteins. Further binding studies, using ligand blot techniques and Superose 12HR fractionation in the presence of [125I]activin, demonstrated UTMP to be an activin-binding protein with a lower affinity for activin than that of follistatin. A study of the specific binding behavior of UTMP to activin, using surface plasmon resonance, revealed an apparent equilibrium dissociation constant (Kd) of 49 +/- 25 nM, compared with the follistatin-activin Kd of 379 +/- 51 pM. Similar to another activin-binding protein, alpha2-macroglobulin, UTMP was unable to neutralize the bioactivity of activin in a bioassay based on the capacity of activin to inhibit the proliferation of an MPC-11 plasmacytoma cell line. The high concentrations of this protein in uterine fluid during pregnancy and its ability to bind activin suggest that UTMP may act as a low affinity, high capacity binding protein to sequester activin in the local uterine environment.
McFarlane
J R
JR
Department of Physiology, University of New England, Armidale, New South Wales, Australia.
Foulds
L M
LM
O'Connor
A E
AE
Phillips
D J
DJ
Jenkin
G
G
Hearn
M T
MT
de Kretser
D M
DM
D028341:Activins; D000482:Allantois / Q000378:metabolism*; D000818:Animals; D001826:Body Fluids / Q000378:metabolism*; D005260:Female; D038681:Follistatin; D006023:Glycoproteins / Q000378:metabolism*; D007265:Inhibins / Q000378:metabolism; D011247:Pregnancy; D015843:Serpins; D012756:Sheep / Q000378:metabolism*
D038681:Follistatin; D006023:Glycoproteins; D015843:Serpins; C029708:uterine luminal fluid proteins; D028341:Activins; D007265:Inhibins