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Molecular cloning and characterization of human kinectin
A Fütterer1, G Kruppa, B Krämer
1Institut für Medizinische Mikrobiologie und Hygiene, Technische Universität München, Germany.
Molecular Biology of the Cell
|February 1, 1995
Summary
Researchers identified human kinectin, a protein homologous to chicken kinectin, which binds to the organelle motor kinesin. This integral membrane protein is anchored in the endoplasmic reticulum, playing a role in intracellular transport.
Area of Science:
- Cell Biology
- Molecular Biology
- Protein Biochemistry
Background:
- Kinesin is a motor protein crucial for intracellular organelle transport.
- The precise receptor for kinesin in the endoplasmic reticulum membrane was previously unidentified.
Purpose of the Study:
- To identify and characterize the human homolog of chicken kinectin.
- To elucidate the cellular localization and function of human kinectin.
Main Methods:
- Human cDNA library screening and hybridization.
- Protein expression analysis (Western blotting, immunoprecipitation).
- Immunofluorescence microscopy using monoclonal antibodies (CT-1).
- Epitope tagging and analysis of protein domains.
Main Results:
- Identified a human cDNA encoding a 156 kDa protein homologous to chicken kinectin.
- Monoclonal antibody CT-1 confirmed evolutionary conservation, showing perinuclear and endoplasmic reticulum-like staining.
- CT-1 and anti-chicken kinectin antibodies cross-reacted with a 160 kDa protein.
- Kinectin possesses a transmembrane domain, anchoring it to the endoplasmic reticulum; nuclear localization sequence is non-functional.
Conclusions:
- Human kinectin is an integral membrane protein of the endoplasmic reticulum.
- Kinectin likely functions as a receptor for the kinesin motor protein.
- Its localization suggests a role in kinesin-mediated intracellular transport processes.