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Isolation and Partial Characterization of CD36 from Skim Milk¹

Department of Food Science, North Carolina State University, Raleigh 27695-7624

Corresponding author:
H. E. Swaisgood; e-mail:
harold_swaisgood{at}ncsu.edu.

CD36, a common milk fat globule membrane glycoprotein, was isolated from skim milk by methods similar to those previously utilized for the isolation of sulfhydryl oxidase. Two separate methods that were employed gave similar purity as observed by electrophoresis. The first was based on differential centrifugation and size-exclusion chromatography, whereas the second combined ultrafiltration and affinity chromatography. After significant purification, the protein was identified by Western blotting and sequence analysis. Deglycosylation decreased the apparent molecular mass from approximately 85 to 57 kDa. These results suggested tissue-specific glycosylation. The purified fractions also exhibited low levels of sulfhydryl oxidase activity, the significance of which will require further study.

Abbreviation key: EDC = 1-ethyl-3- (diethylaminopropyl)carbodiimide, , HABA = 2- (4'-Hydroxybenzeneazo) benzoic acid, , LDL = low-density lipoprotein, , MFGM =milk fat globule membrane, , SOX = sulfhydryl oxidase

Key Words: CD36 • milk fat globule membrane • skim milk • sulfhydryl oxidase