Formation and accumulation of lipolysosomes in developing chick hepatocytes

Formation and accumulation of lipolysosomes in developing chick hepatocytes were investigated by means of electron microscopy in combination with biochemical analyses of the lipid composition in liver homogenates. The lipolysosomes occurred with highest frequency from days 11 to 14 of incubation. They were usually small and electron-dense, but during development they gradually enlarged with an accompanying reduction in electron density. Coinciding with this enlargement was an accumulation of esterified cholesterol in the liver homogenates. After hatching, an immediate decrease in the size and number of lipolysosomes occurred along with a reduction in the concentration of esterified cholesterol, of which only a very small amount remained by 9 days of age. Instead of cholesterol, triglycerides subsequently increased in concentration and accounted for the major lipid content of the liver homogenates. In keeping with the ultrastructural changes, the total volume of cytoplasmic lipid droplets rapidly increased with increasing age. This transient accumulation of esterified cholesterol within lipolysosomes may be attributed to an excessive uptake and processing of plasma lipoprotein particles, probably derived from the egg yolk. This concept is supported by an abundance of coated pits, endosomes and multivesicular bodies in the embryonic hepatocytes.     

Circular dichroic and 1H-NMR studies on the aged form of bovine plasma albumin.

Bovine plasma albumin (BPA) has 17 disulfide bonds and approximately one SH group at Cys-34 which catalyzes the intramolecular SH, S-S exchange reaction in the alkaline region at low ionic strength, resulting in the formation of the aged form (A-form). 1) Fractions of alpha-helix (f alpha) and beta-form (f beta) of iodoacetamide-blocked non-aged BPA (IA-BPA) at pH 6.5 (the N-form) and 9.0 (the B-form) in the absence of added salt were 0.70, 0.12 and 0.62, 0.18, respectively (Era et al. (1990]. However, there were no changes in f alpha and f beta of the iodoacetamide-blocked A-form (IA-A-form) over the pH range from 5.5 to 9.1 in the absence of added salt or in 0.10 M KCl (f alpha approximately 0.60, f beta approximately 0.20), indicating that the secondary structure of the IA-A-form might be similar to that of non-aged IA-BPA at pH 9.0 (B-form) in the absence of added salt, that is, the frozen B-form, stabilized covalently by the repairing of disulfide bonds. 2) The rigidity of the A- and IA-A-forms, as monitored by cross-relaxation times between irradiated and observed protein protons, was similar to or slightly higher than that of non-aged IA-BPA or BMA.(ABSTRACT TRUNCATED AT 250 WORDS)     

Water structure in modified bovine plasma albumin (BPA) gel and bovine mercaptalbumin solution. 1H-n.m.r. studies

Bovine plasma albumin Fr. V (BPA) has been known to contain small amounts of proteolytic enzyme. Wilson & Foster (1971) found a very limited and specific cleavage of BPA catalyzed by the enzyme with BPA in the F-form near pH 3.8, resulting in the formation of partially hydrolyzed BPA (BPA*). BPA* had a tendency to form a transparent gel at pD 4.0 (pD range of the F-form) above 8%, though proteolytic enzyme-free bovine mercaptalbumin (BMA) was in a transparent solution at pD 4.0 even at 12%. Water structures of the F-form of BMA in the solution state and of BPA* in the gel state were studied by measuring 1H-n.m.r. spectra, spin-lattice relaxation time (T1) and cross relaxation time (TIS) between irradiated and observed protons. Protein concentration-dependent changes of T1 of water protons indicated that the amount of hydrated water of BPA* in the gel state is far greater than that of the F-form of BMA in the solution state. TIS values from protein protons to water protons also indicated a large amount of hydration of BPA*, strong interaction between water and BPA* and rapid exchange between bound and bulk water in the gel state.