Research

Our research builds upon 30+ years of continuously funded work on the enzymology and structural biology of proteins involved in blood coagulation and on mechanisms of ligand binding.

Our approach uses a combination of conventional (rapid kinetics, protein engineering, X-ray crystallography, NMR) and state-of-the-art (smFRET, cryo-EM) biophysical approaches that make our lab uniquely positioned to tackle unsolved problems in the field.

Notable contributions of the lab include:

  1. Elucidation of allostery in trypsin-like proteases and their zymogens
  2. Development of a thrombin mutant currently in Phase II testing (NCT03963895)
  3. Elucidation of the general properties of conformational selection and induced fit
  4. Cryo-EM of coagulation factors

Ongoing research includes:

  1. Structure and dynamics of prothrombin (HL049413)
  2. Mechanism of protein C activation (HL139554)
  3. Molecular determinants of thrombin allostery (HL147821)

We are interested in two basic interactions of the blood coagulation cascade, i.e., the interaction of prothrombin with prothrombinase, leading to generation of thrombin and blood clotting, and the interaction of the thrombin-thrombomodulin complex with protein C, initiating the negative feed-back loop that shuts down the coagulation response. Both of these interactions have been studied for decades by several groups but remain in need of structural information.

Our goal is to solve the molecular architecture of prothrombin and protein C, free and bound to their biological activators, and to gain information on the underlying dynamics, mechanism of activation, and epitopes involved in the interactions.

Relevant Bibliography

  1. Serine peptidases: classification, structure and function. Page MJ, Di Cera E (2008) Cell Mol Life Sci 65, 1220-1236.
  2. Allostery in trypsin-like proteases suggests new therapeutic strategies. Gohara DW, Di Cera E (2011) Trends Biotechnol 29, 577-585.
  3. Conformational selection or induced fit? A critical appraisal of the kinetic mechanism. Vogt AD, Di Cera E (2012) Biochemistry 51, 5894-5902.
  4. Mechanisms of ligand binding. Di Cera E (2020) Biophys Rev 1, 011303.
  5. Cryo-EM structures of human coagulation factors V and Va. Ruben EA, Rau MJ, Fitzpatrick JAJ, Di Cera E (2021) Blood, in press.

Complete Bibliography via PubMed: Di Cera E