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Isolation and partial characterization of CD36 from skim milk
C P Wilcox1, V G Janolino, H E Swaisgood
1Department of Food Science, North Carolina State University, Raleigh 27695-7624, USA.
Journal of Dairy Science
|September 7, 2002
Summary
Researchers purified CD36, a milk fat globule membrane glycoprotein, using two methods. The study revealed tissue-specific glycosylation and low sulfhydryl oxidase activity in purified CD36.
Area of Science:
- Biochemistry
- Molecular Biology
- Food Science
Background:
- CD36 is a glycoprotein found in milk fat globule membranes.
- Understanding CD36's properties is crucial for dairy science and nutrition.
- Previous methods exist for isolating related milk proteins like sulfhydryl oxidase.
Purpose of the Study:
- To isolate and characterize CD36 from skim milk.
- To investigate the glycosylation pattern of CD36.
- To assess the enzymatic activity of purified CD36.
Main Methods:
- Isolation of CD36 using differential centrifugation, size-exclusion chromatography, ultrafiltration, and affinity chromatography.
- Protein purity assessment via electrophoresis.
- Identification and characterization using Western blotting and sequence analysis.
- Deglycosylation to analyze molecular mass changes.
Main Results:
- Two distinct purification methods yielded highly pure CD36.
- Deglycosylation reduced the molecular mass from 85 kDa to 57 kDa, indicating tissue-specific glycosylation.
- Purified CD36 demonstrated low levels of sulfhydryl oxidase activity.
Conclusions:
- CD36 can be effectively purified from skim milk using established and novel chromatographic techniques.
- The observed molecular mass shift post-deglycosylation suggests differential glycosylation of CD36 in different tissues.
- The presence of sulfhydryl oxidase activity in purified CD36 warrants further investigation into its functional significance.