Related Experiment Videos
Isolation of plasma membrane complexes from Xenopus oocytes
1Department of Cell Biology and Anatomy, Cornell University Medical College, New York, New York 10021.
The Journal of Membrane Biology
|February 1, 1989
Summary
Researchers developed a new method to isolate Xenopus oocyte plasma membranes, preserving key structures. This technique aids in studying membrane protein sorting and cell surface components.
Area of Science:
- Cell Biology
- Biochemistry
- Developmental Biology
Background:
- Xenopus oocytes are a valuable model for studying cellular processes.
- Isolation of intact plasma membranes is crucial for detailed analysis.
- Previous methods may compromise membrane integrity or composition.
Purpose of the Study:
- To develop and characterize a novel method for isolating plasma membrane fractions from Xenopus oocytes.
- To preserve the morphological integrity of microvilli and coated pits.
- To enable biochemical and morphological studies of membrane protein sorting.
Main Methods:
- Developed a method involving gentle homogenization and Triton X-114 detergent extraction.
- Separated plasma membranes from vitelline envelopes and cortical granules.
- Utilized biochemical assays (5'-nucleotidase, alkaline phosphodiesterase) and lectin techniques (Concanavalin A, wheat germ agglutinin) for characterization.
Main Results:
- Successfully isolated plasma membrane complexes with preserved microvilli and coated pits.
- Achieved substantial recovery (44-66%) of key membrane markers like 5'-nucleotidase and alkaline phosphodiesterase.
- Identified major glycoprotein species using lectin blotting and affinity chromatography.
Conclusions:
- The developed method provides high-quality plasma membrane fractions from Xenopus oocytes.
- These fractions are suitable for detailed biochemical and morphological investigations.
- The technique is particularly useful for studying membrane protein sorting in this model system.