A λ-Dynamics Investigation of Insulin and Other A3 Variant Binding Affinities to the Insulin Receptor.

Insulin is a clinical insulin variant where a conserved valine at the third residue on insulin's A chain (Val) is replaced with a leucine (Leu), weakening insulin receptor (IR) binding by 140-500-fold. This severe impact on binding from a subtle modification has posed an intriguing problem for decades. Although experimental investigations of natural and unnatural A3 mutations have highlighted the sensitivity of insulin-IR binding at this site, atomistic explanations of these binding trends have remained elusive.

A λ-dynamics investigation of insulin and other A3 variant binding affinities to the insulin receptor.

Insulin is a clinical insulin variant where a conserved valine at the third residue on insulin's A chain (Val) is replaced with a leucine (Leu), impairing insulin receptor (IR) binding by 140-500 fold. This severe impact on binding from such a subtle modification has posed an intriguing problem for decades. Although experimental investigations of natural and unnatural A3 mutations have highlighted the sensitivity of insulin-IR binding to minor changes at this site, an atomistic explanation of these binding trends has remained elusive.

Mutant thermal proteome profiling for characterization of missense protein variants and their associated phenotypes within the proteome.

Temperature-sensitive (TS) missense mutants have been foundational for characterization of essential gene function. However, an unbiased approach for analysis of biochemical and biophysical changes in TS missense mutants within the context of their functional proteomes is lacking. We applied MS-based thermal proteome profiling (TPP) to investigate the proteome-wide effects of missense mutations in an application that we refer to as utant hermal roteome rofiling (mTPP).