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Human D-Phe-Pro-Arg-CH2-alpha-thrombin crystallization and diffraction data.

E Skrzypczak-Jankun¹, T J Rydel, A Tulinsky

¹Department of Chemistry, Michigan State University, East Lansing 48824.

Journal of Molecular Biology

|April 20, 1989

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Researchers crystallized human alpha-thrombin inhibited by D-Phe-Pro-Arg-chloromethylketone. This breakthrough enables detailed structural analysis for potential therapeutic development.

Area of Science:

Biochemistry
Structural Biology
Crystallography

Background:

Human alpha-thrombin is a key enzyme in blood coagulation.
Inhibitors are crucial for understanding enzyme function and developing therapeutics.
High-affinity irreversible inhibitors offer specific targeting capabilities.

Purpose of the Study:

To crystallize human alpha-thrombin inhibited by D-Phe-Pro-Arg-chloromethylketone.
To obtain high-quality crystals suitable for X-ray diffraction.
To prepare for a complete crystallographic structure determination.

Main Methods:

Inhibition of human alpha-thrombin with D-Phe-Pro-Arg-chloromethylketone.
Crystallization using polyethylene glycol 8000 and sodium phosphate buffer.

Related Experiment Videos

X-ray diffraction analysis of the obtained crystals.

Main Results:

Successfully crystallized inhibited human alpha-thrombin.
Orthorhombic crystal structure determined with specific lattice parameters (a=67.9(1) A, b=87.9(1) A, c=61.0(1) A).
Space group identified as P2(1)2(1)2(1) with four molecules per unit cell, yielding a 58% protein fraction.

Conclusions:

The obtained crystals exhibit excellent X-ray diffraction quality.
This crystallization is a critical step towards determining the complete 3D structure of inhibited alpha-thrombin.
A mercury heavy-atom derivative is being prepared to facilitate structure determination.