Interaction of locust apolipophorin III with lipoproteins and phospholipid vesicles: effect of glycosylation

• Published in: Journal of lipid research Vol. 41; no. 3; pp. 416 - 423
• Main Authors: Weers, P.M.M, Horst, D.J. van der, Ryan, R.O
• Format: Journal Article
• Language: English
• Published: United States Elsevier 01.03.2000
• Subjects: Animals; apolipoproteins; Apolipoproteins - chemistry; Apolipoproteins - metabolism; apoLp-III; Carbohydrate Conformation; Carbohydrate Sequence; exchangeable apolipoproteins; Glycosylation; Grasshoppers; lipid-protein interactions; lipophorin; Lipoproteins - metabolism; locust; Locusta migratoria; Mass Spectrometry; Molecular Sequence Data; Molecular Structure; Phospholipids - metabolism; Protein Binding; Spectrometry, Fluorescence
• ISSN: 0022-2275; 1539-7262
• DOI: 10.1016/S0022-2275(20)34480-1

Apolipophorin III (apoLp-III) from Locusta migratoria is an exchangeable apolipoprotein that binds reversibly to lipoprotein surfaces. The native protein is glycosylated at Asn-18 and Asn-85. Variable attachment of five distinct oligosaccharide moieties at the two glycosylation sites results in molecular weight heterogeneity, as seen by mass spectrometry. The main mass peak of 20,488 Da decreases to 17,583 Da after removal of carbohydrate, indicating that apoLp-III carbohydrate mass is approximately 14% by weight. Deglycosylated apoLp-III induced clearance of dimyristoylphosphatidylcholine and dimyristoylphosphatidylglycerol vesicles at a faster rate than glycosylated apoLp-III. However, in lipoprotein binding assays, in which apoLp-III interacts with surface-localized diacylglycerol, only minor differences in binding were observed. The fluorescence properties of 1-anilinonaphthalene-8-sulfonate were unaffected by the glycosylation state of apoLp-III, indicating that no changes in the relative amount of exposed hydrophobic surface occurred as a result of carbohydrate removal. We propose that glycosyl moieties affect the ability of apoLp-III to transform phospholipid bilayer vesicles into disc-like complexes by steric hindrance. This is due to the requirement that apoLp-III penetrate the bilayer substrate prior to conformational opening of the helix bundle. On the other hand, the glycosyl moieties do not affect lipoprotein binding interactions as it does not involve deep protein penetration into the lipid milieu. Rather, lipoprotein binding is based on oriented protein contact with the lipid surface followed by opening of the helix bundle, which allows formation of a stable interaction with surface exposed hydrophobic sites.