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Clathrin coated vesicles in Neurospora crassa
1Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC), UNC-CONICET, Argentina.
Molecular and Cellular Biochemistry
|September 1, 1987
Summary
Researchers identified a clathrin-like protein in Neurospora crassa cells. This protein is associated with coated vesicles, essential for intracellular transport, and exhibits properties similar to clathrin found in other organisms.
Area of Science:
- Cell Biology
- Molecular Biology
- Mycology
Background:
- Clathrin is a protein crucial for forming coated vesicles involved in intracellular trafficking.
- The presence and function of clathrin in fungi, such as Neurospora crassa, are not fully understood.
Purpose of the Study:
- To investigate the presence and characteristics of a clathrin-like protein in Neurospora crassa.
- To determine the subcellular localization and assembly properties of this putative protein.
Main Methods:
- Differential centrifugation and subcellular fractionation to isolate cellular components.
- Glass bead permeation chromatography for protein purification.
- Electron microscopy for visualizing coated vesicles.
- Biochemical assays to assess protein association and dissociation.
Main Results:
- A polypeptide with mobility similar to bovine clathrin was detected in N. crassa homogenates.
- The putative clathrin was enriched in the microsomal fraction and further purified.
- Coated vesicles, approximately 42.5 nm in diameter, were identified.
- The protein remained associated with coated vesicles under various conditions but was released by Tris buffer and re-sedimented in Mes buffer, forming structures resembling empty coats.
Conclusions:
- Neurospora crassa possesses a clathrin-like protein involved in the formation of coated vesicles.
- This protein shares functional and structural similarities with clathrin from higher eukaryotes.
- The findings contribute to understanding intracellular transport mechanisms in fungi.