The structural basis of lipid interactions in lipovitellin, a soluble lipoprotein.
BACKGROUND
The conformation and assembly of lipoproteins, protein containing large amounts of noncovalently bound lipid, is poorly understood. Lipoproteins present an unusual challenge as they often contain varying loads of lipid and are not readily crystallized. Lipovitellin is a large crystallizable oocyte protein of approximately 1300 residues that contains about 16% w/w lipid. Lipovitellin contains two large domains that appear to be conserved in both microsomal triglyceride transfer protein and apolipoprotein B-100. To gain insight into the conformation of a lipoprotein and the potential modes of binding of both neutral and phospholipid, the crystal structure of lamprey lipovitellin has been determined.


RESULTS
We report here the refined crystal structure of lipovitellin at 2.8 A resolution. The structure contains 1129 amino acid residues located on five peptide chains, one 40-atom phosphatidylcholine, and one 13-atom hydrocarbon chain. The protein contains a funnel-shaped cavity formed primarily by two beta sheets and lined predominantly by hydrophobic residues.


CONCLUSIONS
Using the crystal structure as a template, a model for the bound lipid is proposed. The lipid-binding cavity is formed primarily by a single-thickness beta-sheet structure which is stabilized by bound lipid. This cavity appears to be flexible, allowing lipid to be loaded or unloaded.
Department of Biochemistry, University of Minnesota, Minneapolis 55455, USA.
T A
TA
Anderson
D G
DG
Levitt
L J
LJ
Banaszak
D000595:Amino Acid Sequence; D001055:Apolipoproteins B / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism; D018360:Crystallography, X-Ray; D004527:Egg Proteins; D015387:Egg Proteins, Dietary; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000378:metabolism; D011487:Protein Conformation; D015032:Zinc / Q000378:metabolism
D001055:Apolipoproteins B; D002352:Carrier Proteins; D004527:Egg Proteins; D015387:Egg Proteins, Dietary; D008055:Lipids; D010743:Phospholipids; C106092:microsomal triglyceride transfer protein; C001549:lipovitellin; D015032:Zinc