دورية
Arginine 200 of Heparin Cofactor II Promotes Intramolecular Interactions of the Acidic Domain
| العنوان: | Arginine 200 of Heparin Cofactor II Promotes Intramolecular Interactions of the Acidic Domain |
|---|---|
| المؤلفون: | Ciaccia, Angelina V., Monroe, Dougald M., Church, Frank C. |
| المصدر: | Journal of Biological Chemistry; May 1997, Vol. 272 Issue: 22 p14074-14079, 6p |
| مستخلص: | Heparin cofactor II (HCII) is presumed to be a physiological inhibitor of the serine proteinase thrombin. The reaction between HCII and thrombin is quite unique, because it involves an unusual HCII-reactive site loop sequence of Leu444-Ser445, requires the presence of glycosaminoglycans for optimal activity and involves a protein-protein interaction besides the reactive site loop-active site interaction characteristic of serine proteinase inhibitor-serine proteinase pairs. Two mutations at a unique HCII residue, Arg200→ Ala or Glu, were generated by site-directed mutagenesis. The mutations did not alter either HCII binding to heparin-Sepharose or HCII inhibition of thrombin in the presence of heparin or dermatan sulfate, suggesting that Arg200is not part of the glycosaminoglycan binding site of HCII. In the absence of glycosaminoglycan, there was a significant increase in α-thrombin inhibition by the Arg200mutants as compared with wild type recombinant HCII (wt-rHCII), whereas inhibition rates with chymotrypsin were identical. Inhibition of γT-thrombin, which lacks anion-binding exosite 1 ((ABE-1), the region of α-thrombin that interacts with the acidic domain of HCII), was significantly reduced compared with α-thrombin, but the reduction was more dramatic for the Arg200-rHCII mutants. Hirugen, which binds to ABE-1 of α-thrombin, also diminished inhibition of α-thrombin by the Arg200-rHCII mutants to nearly wt-rHCII levels. Both Arg200-rHCII mutants had significantly increasedkavalues as compared with wt-rHCII, whereas thekdrates were unchanged. Collectively, these results suggest that the improved inhibitory activity of the Arg200-rHCII mutants is mediated by enhanced interactions between the acidic domain and ABE-1, resulting in an increased HCII-thrombin association rate. |
| قاعدة البيانات: | Supplemental Index |
| تدمد: | 00219258 1083351X |
|---|---|
| DOI: | 10.1074/jbc.272.22.14074 |