The allosteric landscape of the Src kinase

Area of Science:

• Biochemistry and enzymology
• Structural biology
• Drug discovery

Background:

• Enzymes are crucial biological catalysts and drug targets.
• Current drugs often target active sites, leading to specificity issues.
• Allosteric sites offer greater specificity but are largely unmapped in enzymes.

Purpose of the Study:

• To create a comprehensive experimental allosteric map of the Src protein kinase.
• To understand the nature and extent of allosteric regulation in kinases.
• To identify novel druggable allosteric sites on Src kinase.

Main Methods:

• Quantified effects of over 50,000 amino acid substitutions on enzyme activity and abundance.
• Employed thermodynamic modeling to differentiate fold stability and catalytic changes.
• Developed a comprehensive energy landscape of the kinase domain.

Main Results:

• Revealed extensive, directionally biased allosteric networks across the Src kinase domain.
• Demonstrated distance-dependent decay of inhibitory allosteric effects from the active site.
• Identified previously unreported, potentially druggable allosteric sites on Src.

Conclusions:

• Established a framework for comprehensive allosteric mapping in kinases and other enzymes.
• The Src allosteric map provides insights into enzyme regulation and drug development.
• Allosteric site targeting can enhance drug specificity and reduce toxicity.