Combinatorial and Computational Approaches to Identify Interactions of Macrophage Colony-stimulating Factor (M-CSF) and Its Receptor c-FMS

Citation:

L. Rosenfeld, J. Shirian, Y. Zur, N. Levaot, J. M. Shifman, and N. Papo. 2015. “Combinatorial and Computational Approaches to Identify Interactions of Macrophage Colony-stimulating Factor (M-CSF) and Its Receptor c-FMS.” J Biol Chem, 290, Pp. 26180-93.

Abstract:

The molecular interactions between macrophage colony-stimulating factor (M-CSF) and the tyrosine kinase receptor c-FMS play a key role in the immune response, bone metabolism, and the development of some cancers. Because no x-ray structure is available for the human M-CSF . c-FMS complex, the binding epitope for this complex is largely unknown. Our goal was to identify the residues that are essential for binding of the human M-CSF to c-FMS. For this purpose, we used a yeast surface display (YSD) approach. We expressed a combinatorial library of monomeric M-CSF (M-CSFM) single mutants and screened this library to isolate variants with reduced affinity for c-FMS using FACS. Sequencing yielded a number of single M-CSFM variants with mutations both in the direct binding interface and distant from the binding site. In addition, we used computational modeling to map the identified mutations onto the M-CSFM structure and to classify the mutations into three groups as follows: those that significantly decrease protein stability; those that destroy favorable intermolecular interactions; and those that decrease affinity through allosteric effects. To validate the YSD and computational data, M-CSFM and three variants were produced as soluble proteins; their affinity and structure were analyzed; and very good correlations with both YSD data and computational predictions were obtained. By identifying the M-CSFM residues critical for M-CSF . c-FMS interactions, we have laid down the basis for a deeper understanding of the M-CSF . c-FMS signaling mechanism and for the development of target-specific therapeutic agents with the ability to sterically occlude the M-CSF.c-FMS binding interface.

Notes:

Rosenfeld, LiorShirian, JasonZur, YuvalLevaot, NoamShifman, Julia MPapo, NivengResearch Support, Non-U.S. Gov't2015/09/12 06:00J Biol Chem. 2015 Oct 23;290(43):26180-93. doi: 10.1074/jbc.M115.671271. Epub 2015 Sep 10.