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Structure of the hirugen and hirulog 1 complexes of alpha-thrombin.
J Mol Biol. 1991 Oct 20; 221(4):1379-93.JM

Abstract

The isomorphous structures of the hirugen (N-acetylhirudin 53'-64' with sulfato-Tyr63') and hirulog 1 (D-Phe-Pro-Arg-Pro-(Gly)4 desulfato-Tyr63'-hirugen) complexes of human alpha-thrombin have been determined and refined at 2.2 A resolution to crystallographic R-factors of 0.167 and 0.163, respectively. The binding of hirugen to thrombin is similar to that of the binding of the C-terminal dodecapeptide of hirudin, including that of the terminal 3(10) helical turn. The sulfato Tyr63', which, as a result of sulfation, increases the binding affinity by an order of magnitude, is involved in an extended hydrogen bonding network utilizing all three sulfato oxygen atoms. The hirugen-thrombin complex is the first thrombin structure determined to have an unobstructed active site; this site is practically identical in positioning of catalytic residues and in its hydrogen bonding pattern with that of other serine proteinases. Hirulog 1, which is a poor thrombin substrate, is cleaved at the Arg3'-Pro4' bond in the crystal structure. The Arg3' of hirulog 1 occupies the specificity site, the D-Phe-Pro-Arg tripeptide is positioned like that of D-Phe-Pro-Arg chloromethylketone in the active site and the Pro4'(Gly)4 spacer to hirugen is disordered in the structure, as is the 3(10) turn of hirugen. The latter must be related to the simultaneous absence both of sulfation and of the last residue of hirudin (Gln65'). In addition, the autolysis loop of thrombin (Lys145-Gly150) is disordered in both structures. Changes in circular dichroism upon hirugen binding are therefore most likely the result of the flexibility associated with this loop.

Authors+Show Affiliations

Department of Chemistry, Michigan State University, East Lansing 48824-1322.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

1942057

Citation

Skrzypczak-Jankun, E, et al. "Structure of the Hirugen and Hirulog 1 Complexes of Alpha-thrombin." Journal of Molecular Biology, vol. 221, no. 4, 1991, pp. 1379-93.
Skrzypczak-Jankun E, Carperos VE, Ravichandran KG, et al. Structure of the hirugen and hirulog 1 complexes of alpha-thrombin. J Mol Biol. 1991;221(4):1379-93.
Skrzypczak-Jankun, E., Carperos, V. E., Ravichandran, K. G., Tulinsky, A., Westbrook, M., & Maraganore, J. M. (1991). Structure of the hirugen and hirulog 1 complexes of alpha-thrombin. Journal of Molecular Biology, 221(4), 1379-93.
Skrzypczak-Jankun E, et al. Structure of the Hirugen and Hirulog 1 Complexes of Alpha-thrombin. J Mol Biol. 1991 Oct 20;221(4):1379-93. PubMed PMID: 1942057.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Structure of the hirugen and hirulog 1 complexes of alpha-thrombin. AU - Skrzypczak-Jankun,E, AU - Carperos,V E, AU - Ravichandran,K G, AU - Tulinsky,A, AU - Westbrook,M, AU - Maraganore,J M, PY - 1991/10/20/pubmed PY - 1991/10/20/medline PY - 1991/10/20/entrez SP - 1379 EP - 93 JF - Journal of molecular biology JO - J Mol Biol VL - 221 IS - 4 N2 - The isomorphous structures of the hirugen (N-acetylhirudin 53'-64' with sulfato-Tyr63') and hirulog 1 (D-Phe-Pro-Arg-Pro-(Gly)4 desulfato-Tyr63'-hirugen) complexes of human alpha-thrombin have been determined and refined at 2.2 A resolution to crystallographic R-factors of 0.167 and 0.163, respectively. The binding of hirugen to thrombin is similar to that of the binding of the C-terminal dodecapeptide of hirudin, including that of the terminal 3(10) helical turn. The sulfato Tyr63', which, as a result of sulfation, increases the binding affinity by an order of magnitude, is involved in an extended hydrogen bonding network utilizing all three sulfato oxygen atoms. The hirugen-thrombin complex is the first thrombin structure determined to have an unobstructed active site; this site is practically identical in positioning of catalytic residues and in its hydrogen bonding pattern with that of other serine proteinases. Hirulog 1, which is a poor thrombin substrate, is cleaved at the Arg3'-Pro4' bond in the crystal structure. The Arg3' of hirulog 1 occupies the specificity site, the D-Phe-Pro-Arg tripeptide is positioned like that of D-Phe-Pro-Arg chloromethylketone in the active site and the Pro4'(Gly)4 spacer to hirugen is disordered in the structure, as is the 3(10) turn of hirugen. The latter must be related to the simultaneous absence both of sulfation and of the last residue of hirudin (Gln65'). In addition, the autolysis loop of thrombin (Lys145-Gly150) is disordered in both structures. Changes in circular dichroism upon hirugen binding are therefore most likely the result of the flexibility associated with this loop. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/1942057/Structure_of_the_hirugen_and_hirulog_1_complexes_of_alpha_thrombin_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0022-2836(91)90939-4 DB - PRIME DP - Unbound Medicine ER -