Reformable intramolecular cross-linking of the N-terminal domain of heparin cofactor II - Effects on enzyme inhibition

Brinkmeyer S, Eckert R, Ragg H (2004)
EUROPEAN JOURNAL OF BIOCHEMISTRY 271(21): 4275-4283.

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Abstract
The crystal structure of a heparin cofactor II (HCII)-thrombin Michaelis complex has revealed extensive contacts encompassing the N-terminal domain of HCII and exosite I of the proteinase. In contrast, the location of the N-terminal extension in the uncomplexed inhibitor was unclear. Using a disulfide cross-linking strategy, we demonstrate that at least three different sites (positions 52, 54 and 68) within the N terminus may be tethered in a reformable manner to position 195 in the loop region between helix D and strand s2A of the HCII molecule, suggesting that the N-terminal domain may interact with the inhibitor scaffold in a permissive manner. Cross-linking of the N terminus to the HCII body does not strongly affect the inhibition of alpha-chymotrypsin, indicating that the reactive site loop sequences of the engineered inhibitor variants, required for interaction with one of the HCII target enzymes, are normally accessible. In contrast, intramolecular tethering of the N-terminal extension results in a drastic decrease of alpha-thrombin inhibitory activity, both in the presence and in the absence of glycosaminoglycans. Treatment with dithiothreitol and iodoacetamide restores activity towards alpha-thrombin, suggesting that release of the N terminus of HCII is an important component of the multistep interaction between the inhibitor and alpha-thrombin.
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Brinkmeyer S, Eckert R, Ragg H. Reformable intramolecular cross-linking of the N-terminal domain of heparin cofactor II - Effects on enzyme inhibition. EUROPEAN JOURNAL OF BIOCHEMISTRY. 2004;271(21):4275-4283.
Brinkmeyer, S., Eckert, R., & Ragg, H. (2004). Reformable intramolecular cross-linking of the N-terminal domain of heparin cofactor II - Effects on enzyme inhibition. EUROPEAN JOURNAL OF BIOCHEMISTRY, 271(21), 4275-4283.
Brinkmeyer, S., Eckert, R., and Ragg, H. (2004). Reformable intramolecular cross-linking of the N-terminal domain of heparin cofactor II - Effects on enzyme inhibition. EUROPEAN JOURNAL OF BIOCHEMISTRY 271, 4275-4283.
Brinkmeyer, S., Eckert, R., & Ragg, H., 2004. Reformable intramolecular cross-linking of the N-terminal domain of heparin cofactor II - Effects on enzyme inhibition. EUROPEAN JOURNAL OF BIOCHEMISTRY, 271(21), p 4275-4283.
S. Brinkmeyer, R. Eckert, and H. Ragg, “Reformable intramolecular cross-linking of the N-terminal domain of heparin cofactor II - Effects on enzyme inhibition”, EUROPEAN JOURNAL OF BIOCHEMISTRY, vol. 271, 2004, pp. 4275-4283.
Brinkmeyer, S., Eckert, R., Ragg, H.: Reformable intramolecular cross-linking of the N-terminal domain of heparin cofactor II - Effects on enzyme inhibition. EUROPEAN JOURNAL OF BIOCHEMISTRY. 271, 4275-4283 (2004).
Brinkmeyer, Stephan, Eckert, Ralf, and Ragg, Hermann. “Reformable intramolecular cross-linking of the N-terminal domain of heparin cofactor II - Effects on enzyme inhibition”. EUROPEAN JOURNAL OF BIOCHEMISTRY 271.21 (2004): 4275-4283.
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42 References

Data provided by Europe PubMed Central.

Serpin structure, mechanism, and function.
Gettins PG., Chem. Rev. 102(12), 2002
PMID: 12475206
What does it mean to be natively unfolded?
Uversky VN., Eur. J. Biochem. 269(1), 2002
PMID: 11784292
Stabilization of functional proteins by introduction of multiple disulfide bonds.
Matsumura M, Matthews BW., Meth. Enzymol. 202(), 1991
PMID: 1784181
Active-site titration of peptidases.
Knight CG., Meth. Enzymol. 248(), 1995
PMID: 7674964
Inhibition of chymotrypsin by heparin cofactor II.
Church FC, Noyes CM, Griffith MJ., Proc. Natl. Acad. Sci. U.S.A. 82(19), 1985
PMID: 3863104
A rapid and efficient one-tube PCR-based mutagenesis technique using Pfu DNA polymerase.
Picard V, Ersdal-Badju E, Lu A, Bock SC., Nucleic Acids Res. 22(13), 1994
PMID: 8041621
A new member of the plasma protease inhibitor gene family.
Ragg H., Nucleic Acids Res. 14(2), 1986
PMID: 3003690

Kellner, 1994
Probing protein folding and stability using disulfide bonds.
Darby N, Creighton TE., Mol. Biotechnol. 7(1), 1997
PMID: 9163722

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