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Phosphatidylserine specific binding protein in rat brain: purification and characterization
T Nakaoka1, N Kojima, T Hamamoto
1Glyco Molecular Biology, Frontier Research Program, Institute of Physical and Chemical Research (RIKEN), Saitama.
Journal of Biochemistry
|October 1, 1993
Summary
Myristoylated, alanine-rich C kinase substrate (MARCKS) binds phosphatidylserine in a calcium-independent manner. Phosphorylation by protein kinase C disrupts this specific phosphatidylserine binding, suggesting MARCKS targets this membrane molecule.
Area of Science:
- Biochemistry
- Molecular Biology
- Neuroscience
Background:
- Myristoylated, alanine-rich C kinase substrate (MARCKS) is a protein involved in cellular processes.
- The specific binding partners and membrane interactions of MARCKS are not fully elucidated.
Purpose of the Study:
- To identify and characterize the phosphatidylserine binding protein from rat brain.
- To investigate the role of phosphorylation and myristoylation in MARCKS-phosphatidylserine interactions.
Main Methods:
- Liposome blotting analysis for protein purification.
- Biochemical assays to determine binding specificity.
- Bacterial expression of MARCKS to assess myristoylation role.
Main Results:
- A protein identical to MARCKS was purified and shown to bind phosphatidylserine specifically.
- MARCKS binding to phosphatidylserine was calcium-independent.
- Protein kinase C-dependent phosphorylation abolished MARCKS binding to phosphatidylserine.
- Bacterially expressed MARCKS retained phosphatidylserine binding, indicating myristoylation is not essential.
Conclusions:
- Phosphatidylserine is identified as a membranous target molecule of MARCKS.
- MARCKS exhibits specific binding to phosphatidylserine, which is regulated by phosphorylation.
- The N-terminal myristoylation of MARCKS is not required for phosphatidylserine binding.