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Iodinated fatty acids as probes for myristate processing and function. Incorporation into pp60v-src
S M Peseckis1, I Deichaite, M D Resh
1Department of Cell Biology and Genetics, Memorial Sloan-Kettering Cancer Center, New York, New York 10021.
The Journal of Biological Chemistry
|March 5, 1993
Summary
Researchers synthesized iodinated fatty acids to study myristylated proteins, finding that variations in the myristyl group still allow membrane association for pp60v-src protein.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- The myristyl group is crucial for protein processing, trafficking, and function.
- Myristoylated proteins, like pp60v-src, play key roles in cellular processes.
- Understanding the myristyl group's role in membrane association is vital.
Purpose of the Study:
- To investigate the contribution of the myristyl group to the membrane association of pp60v-src.
- To synthesize and utilize [omega-125I]iodo-fatty acids as analogs of myristate.
- To determine if modified fatty acids can be incorporated into pp60v-src and affect its membrane binding.
Main Methods:
- In vitro translation of v-src mRNA to incorporate fatty acid analogs.
- One-dimensional peptide analysis to confirm analog attachment site.
- In vivo labeling of cells with iodinated fatty acids and subsequent cell fractionation.
Main Results:
- 12-Iodododecanoic, 13-iodotridecanoic, and 14-iodotetradecanoic acids were incorporated into pp60v-src in vitro and in vivo.
- Analog incorporation occurred at the N-terminus of pp60v-src.
- Modified pp60v-src proteins showed comparable or slightly reduced membrane association compared to the myristate-modified protein.
Conclusions:
- Fatty acyl groups differing by one carbon from myristate can be accommodated in the pp60v-src membrane binding site.
- In vitro systems can effectively predict analog behavior in vivo.
- Iodinated fatty acids are valuable tools for studying the function of fatty acids in myristylated molecules.