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Developmentally regulated lectin from embryonic chick pectoral muscle. Purification by affinity chromatography
The Journal of Biological Chemistry
|September 10, 1977
Summary
Researchers purified a muscle lectin from embryonic chick pectoral muscle, finding its activity increases during development. This lectin is located on and within myoblasts, suggesting a role in muscle development.
Area of Science:
- Developmental Biology
- Biochemistry
- Cell Biology
Background:
- Specific lectin activity in embryonic chick pectoral muscle extracts increases significantly between 8 and 16 days of development.
- Lectins are proteins known for their carbohydrate-binding properties, often involved in cellular recognition and adhesion.
Purpose of the Study:
- To purify and characterize a lectin from embryonic chick pectoral muscle.
- To investigate the developmental changes in lectin activity.
- To determine the cellular localization of the purified lectin.
Main Methods:
- Affinity chromatography using p-aminophenyl-beta-D-lactoside coupled to Sepharose 4B for lectin purification.
- Preparative isoelectric focusing to remove contaminating proteins.
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for molecular weight determination.
- Antibody production and immunofluorescence studies for cellular localization.
Main Results:
- A pure lectin was obtained with an apparent molecular weight of 30,000 and subunit molecular weight of 15,000.
- The lectin's isoelectric point was determined to be 4.0.
- Thiodigalactoside and lactose were identified as potent saccharide inhibitors.
- Immunological studies confirmed the lectin's presence on the surface and within myoblasts.
Conclusions:
- A specific lectin involved in embryonic chick muscle development has been successfully purified and characterized.
- The lectin's localization suggests a potential role in myoblast function, possibly in cell-cell interactions or intracellular processes during muscle formation.