2012
DOI: 10.1016/j.ab.2011.11.021
|View full text |Cite
|
Sign up to set email alerts
|

Fluorescent reporters of thrombin, heparin cofactor II, and heparin binding in a ternary complex

Abstract: Thrombin inactivation by heparin cofactor II (HCII) is accelerated by ternary complex formation with heparin. The novel active-site-labeled thrombins, [4′F]FPR-T and [6F]FFR-T, and the exosite I probe, Hir-(54–65)( SO3−), characterized thrombin exosite I and II interactions with HCII and heparin in the complex. HCII binding to exosite I of heparin-bound [4′F]FPR-T caused a saturable fluorescence increase, absent with antithrombin. Heparin binding to exosite II and a second weaker site caused fluorescence quenc… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
1
1

Citation Types

0
4
0

Year Published

2017
2017
2021
2021

Publication Types

Select...
4

Relationship

0
4

Authors

Journals

citations
Cited by 4 publications
(4 citation statements)
references
References 58 publications
0
4
0
Order By: Relevance
“…Structural studies using X-ray crystallography, mass spectrometry, and nuclear magnetic resonance have confirmed inter-exosite allostery, 8,15−18 as have dual ligand binding studies. 14,[19][20][21][22][23]52 Although one group initially failed to observe direct communication between the exosites of thrombin, 24,25 they subsequently reported a reduced level of binding of hirudin 54−65 peptide to exosite 1 of prethrombin 2 in the presence of prothrombin fragment 1.2, which binds exosite 2. 60 These discrepancies could be the result of ligand-specific interactions, as demonstrated by inter-exosite communication mediated by PAR-1 peptide, but not by hirudin 54−65 peptide.…”
Section: ■ Discussionmentioning
confidence: 99%
See 2 more Smart Citations
“…Structural studies using X-ray crystallography, mass spectrometry, and nuclear magnetic resonance have confirmed inter-exosite allostery, 8,15−18 as have dual ligand binding studies. 14,[19][20][21][22][23]52 Although one group initially failed to observe direct communication between the exosites of thrombin, 24,25 they subsequently reported a reduced level of binding of hirudin 54−65 peptide to exosite 1 of prethrombin 2 in the presence of prothrombin fragment 1.2, which binds exosite 2. 60 These discrepancies could be the result of ligand-specific interactions, as demonstrated by inter-exosite communication mediated by PAR-1 peptide, but not by hirudin 54−65 peptide.…”
Section: ■ Discussionmentioning
confidence: 99%
“…Thus, binding of thrombomodulin to exosite 1 or binding of prothrombin fragment 2 to exosite 2 exerts functional effects on the active site. Furthermore, these allosteric effects have been shown to extend to the other exosite, indicative of the global response of thrombin to ligand binding. Thus, binding of peptide or aptamer ligands to one exosite on thrombin exerts functional effects at the other exosite. Although disputed with small synthetic ligands, , inter-exosite allostery has been shown to occur with physiological ligands such as fibrin, where thrombin binding via exosite 1 is attenuated by exosite 2 ligands . Thus, to examine the functional consequences of this interaction, we investigated whether exosite 2-specific ligands influence binding of thrombin to thrombomodulin and protein C activation by the thrombin–thrombomodulin complex.…”
mentioning
confidence: 99%
See 1 more Smart Citation
“…For example, a specific complexity of CD3 bsAbs is efficacy and on‐target toxicity are driven by trimer formation between the bsAb, T cell, and tumor cell 13 . A bell‐shaped concentration vs. response relationship can be observed, which is a well‐described phenomenon for ternary complexes 30–32 . When bsAb concentrations are low, conditions favor bivalent binding and the formation of trimers.…”
Section: Early Clinical Pharmacology Challenges For Bispecific Antibomentioning
confidence: 99%