A. Mansouri et al., RETINOL FREE AND RETINOL COMPLEXED BETA-LACTOGLOBULIN BINDING-SITES IN BOVINE GERM-CELLS, Biochimica et biophysica acta. Molecular cell research, 1357(1), 1997, pp. 107-114
A high affinity specific binding site for bovine beta-lactoglobulin (B
LG) was identified in bovine germ cell plasma membrane enriched fracti
ons. Binding was found to be reversible and pi-I-dependent with maximu
m binding occurring at pH 5. The on-rate and off-rate constants were 2
.26 +/- 0.8 x 10(5) M-1 min(-1) (n = 3) and 0.016 +/- 0.004 min(-1) (n
= 3), respectively. Scatchard analysis showed a single class of bindi
ng sites, with 12.38 +/- 4.62 x 10(12) sites per mg of membrane protei
n (n = 3) and a dissociation constant (K-D) estimated at 26.43 +/- 2.6
8 nM. There was inhibition of iodinated-BLG (variant A) (I-125-BLGA) b
inding to germ cell plasma membrane enriched fractions in the presence
of unlabelled BLG variant A, BLG variant B, retinol complexed BLGA an
d human retinol-binding protein. Inhibition was observed neither with
BSA nor with lactoferrin. I-125-BLGA incubated with a Triton X-100 sol
ubilized plasma membrane fraction formed a high molecular mass complex
in Superose 12B gel filtration. This receptor complex disappeared in
the presence of unlabelled BLGA and in the presence of 10 mM EDTA. The
results suggest that germ cell plasma membrane may contain a receptor
which is capable of binding either retinol free or retinol complexed
BLGA.