Peripheral synthesis and isoform distribution of dog apoprotein E. An in vivo approach.
Purified dog plasma apo-E is composed of four major isoforms with the following pI values: 5.40, 5.31, 5.26, and 5.22. Treatment with neuraminidase suggests that the multiple forms are due to progressive sialation. The acidic isoforms (pI = 5.22 and less), rarely detectable in plasma lipoprotein samples (except in the d less than 1.05 g/ml fraction of cholesterol-fed dogs), are present in high concentrations in the interstitial fluid high density lipoprotein fraction I (HDLI) of cholesterol-fed dogs, a lipoprotein recently described (Dory, L., Boquet, L.M., Hamilton, R.L., Sloop, C.H., and Roheim, P.S. (1985) J. Lipid Res. 26, 519-527). Apo-Es0 (the most basic isoform) is a major constituent of the plasma d less than 1.05 g/ml fraction, but it is usually only a minor component of other plasma or interstitial fluid lipoproteins. This is likely a reflection of the prolonged residence time of a specific lipoprotein species in this density range, resulting in more extensive desialation. Peripheral apo-E synthesis has been measured under in vivo conditions using a hindlimb cannulation of the lymphatics and collection of prenodal peripheral lymph. Following injection of [3H]leucine or [35S]methionine into the dorsal skin of the toes, the specific activity of the interstitial fluid apo-E far exceeded that of plasma apo-E at all time points examined. Incorporation was measurable 30 min after the isotope injection and peaked at 150 min in control dogs and between 120-150 min in cholesterol-fed dogs. The rate of peripheral synthesis of apo-E in cholesterol-fed dogs appeared to be twice that of control dogs. The newly synthesized and secreted apo-E preferentially associated with the disc-shaped HDLI of the interstitial fluid; less than 15% of the apo-E-associated radioactivity was recovered in the d less than 1.05 g/ml fraction, despite the fact that this fraction contains well over 50% of the total interstitial fluid apo-E mass. The newly secreted, HDLI-associated apo-E can be converted by neuraminidase into apo-Es0.
D000818:Animals; D001057:Apolipoproteins E / Q000096:biosynthesis*; D001345:Autoradiography; D002791:Cholesterol, Dietary / Q000494:pharmacology; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D007526:Isoelectric Point; D007930:Leucine / Q000378:metabolism; D013997:Time Factors
D001057:Apolipoproteins E; D002791:Cholesterol, Dietary; D007930:Leucine