Skip to Content
MilliporeSigma
  • Quantitative analysis of protease recognition by inhibitors in plasma using microscale thermophoresis.

Quantitative analysis of protease recognition by inhibitors in plasma using microscale thermophoresis.

Scientific reports (2016-10-16)
T Dau, E V Edeleva, S A I Seidel, R A Stockley, D Braun, D E Jenne
ABSTRACT

High abundance proteins like protease inhibitors of plasma display a multitude of interactions in natural environments. Quantitative analysis of such interactions in vivo is essential to study diseases, but have not been forthcoming, as most methods cannot be directly applied in a complex biological environment. Here, we report a quantitative microscale thermophoresis assay capable of deciphering functional deviations from in vitro inhibition data by combining concentration and affinity measurements. We obtained stable measurement signals for the substrate-like interaction of the disease relevant inhibitor α-1-antitrypsin (AAT) Z-variant with catalytically inactive elastase. The signal differentiates between healthy and sick AAT-deficient individuals suggesting that affinity between AAT and elastase is strongly modulated by so-far overlooked additional binding partners from the plasma.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Elafin human, >90% (by MS, HPLC and SDS-PAGE), recombinant, expressed in Saccharomyces cerevisiae