The 3D structure-function group, within the department of Biochemistry at the Cardiovascular Research Institute Maastricht (Maastricht University), a small and newly started unit.
The
group has a broad interest in the structure-function studies of coagulation
proteins that are involved in homeostasis as well as in many pathological
conditions such as thromboembolism and bleeding.
Focus
Building
on the vast work that has been done by (former) members of the department such as Prof. Coen Hemker, Prof. Rob Zwaal, Prof. Jan Rosing, Dr. Guido Tans, Dr. Theo Lindhout, Dr. Edouard Bevers and Dr. George Willems, we
focuss on the structure-function relationships of
a number of enzymatic reactions that are of utmost importance for the
maintenance of the haemostatic balance. We employ state-of-the-art techniques that provide us with research possibilities that were not available before. The proteins that we focus on mainly
are:
1. coagulation
factor V (FV)
and its regulation by the anticoagulant protein C system. FV is the essential
cofactor of the prothrombin converting prothrombinase complex. In the absence of
FV activity, patients can suffer from life threatening bleedings, whereas
failure to down regulate FV activity may lead to thromboembolic disease (e.g. in
case of FVLeiden).
2. Protein
C,
its formation and structure function relationships. Protein C is the main
protein involved enzymatic control of regulation. Whereas a great excess of
serine protease inhibitors circulates in plasma (e.g. antithrombin or
alpha-2-macroglobulin), a deficiency in protein C may lead to a serious
thrombotic complications.
3.
Regulation
of coagulation factor VIIIa (FVIIIa)
activity by the protein C system. The proteolytic regulation of the activity of
the “tenase” complex by activated protein C is brought about by the interplay
between FVIIIa and APC in a membrane bound complex that employs protein S and FV
as cofactors. It is assumed that e.g. the failure of FVLeiden to express its
anticoagulant cofactor properties is one of the factors that contribute to the
thrombotic phenotype in FVLeiden carriers.
Besides
having our main focus on basic coagulation research, we have an active
participation in several related topics, through collaborations with fellow
research groups. Where possible, we support the patient-oriented research in the
Biochemistry department and in the Internal Medicine department. Furthermore, we
contribute to the development and improval of the diagnostic repertoire in the
field of haemostasis and thrombosis.
Lastly, we contribute to the education of medical students at the Faculty of Health and Medical Life Sciences and provide expert knowledge in biochemistry in general, and structural bioinformatics in particular.
Approach
To
gather new information in the areas described above, we employ various
techniques to the benefit of our research. Techniques that are applied are
amongst others:
Biochemical/Molecular
Biological techniques
Human
and bovine plasma protein purification
protein
characterization (SDS-PAGE, FPLC, HPLC, MS, Immunological techniques,
SPR)
recombinant
protein expression and purification (pro- and eukaryotic
systems)
Enzyme
Kinetics analysis
Fluorescence
measurements/FRET
Structural
Bioinformatics techniques
Homology
modeling and threading
Structure
analysis
Protein-ligand
docking
Molecular
Dynamics simulations
Virtual
Ligand screening
Central
to our research is the information
exchange between experimental (i.e. basic biochemical/molecular
biology/peptide chemistry/surface plasmon resonance) and computational
approaches (docking/molecular modeling/virtual ligand screening) such that new
information on the function of these essential proteins can be obtained. Ideally
we use or propose a molecular model that can explain a reaction mechanism and on
basis of this model we develop a working hypothesis that is tested by means of
mutagenesis study or any other type of study which allows us to proof (or
disproof) our hypotheses.
With
any new information obtained, we can improof our 3D models and thus continue in
the research cycle given below. Ultimately our approach should lead to improved
treatment regimes, diagnostics as well as novel therapeutics in the
cardiovascular field.
To
accomplish our goals, we have a number of greatly valued collaborations with
other expert laboratories .