J. Biol. Chem., Vol. 261, Issue 2, 563-568, Jan, 1986
Lipoprotein interconversions in an insect, Manduca sexta. Evidence for a lipid transfer factor in the hemolymph
RO Ryan, SV Prasad, EJ Henriksen, MA Wells and JH Law
Hemolymph lipoproteins (lipophorins) of adult Manduca sexta are disinct
from larval forms in density, lipid content, composition, and the presence
of a third, low molecular weight apoprotein. Generally, only one
lipoprotein species exists in M. sexta hemolymph during any given life
stage. Progression through the life cycle results in alterations of
existing lipoproteins to produce new forms, without new protein synthesis.
The observed alterations in lipoprotein density could result from
facilitated lipid transfer in insect hemolymph. An in vitro assay of
facilitated lipid transfer was developed which employs a high density
lipophorin from the wandering larva (density = 1.18 g/ml) as acceptor and
adult low density lipophorin (density = 1.03 g/ml) as donor. Adult
lipophorin-deficient hemolymph was shown to catalyze a time-dependent
equilibration of the starting lipoproteins to produce a new intermediate
lipophorin, Lp-I. Hydrodynamic experiments on the donor, acceptor, and
product lipoproteins excluded fusion as the mechanism whereby Lp-I is
produced. Thus, it is concluded that Lp-I results from facilitated net
lipid transfer from low to high density lipoprotein. Furthermore,
experiments conducted with radioiodinated donor and radioiodinated acceptor
lipoproteins demonstrated that apoprotein exchange does not occur during
the lipid transfer reaction. When donor lipoprotein was labeled in the
lipid moiety with carbon-14, evidence of diacylglycerol and phospholipid
exchange was obtained. Partial characterization of the lipid transfer
factor revealed a relationship between incubation time, donor
concentration, acceptor concentration, lipophorin-deficient hemolymph
concentration, and transfer activity, as measured by Lp-I production. It is
concluded that lipophorin-deficient hemolymph contains one or more
factor(s) that catalyze net lipid transfer as well as diacylglycerol and
phospholipid exchange between lipophorins to produce a single form at
equilibrium.