RNA aptamer to thrombin binds anion-binding exosite-2 and alters protease inhibition by heparin-binding serpins

Martha L. Jeter; Linda V. Ly; Yolanda M. Fortenberry; Herbert C. Whinna; Rebekah R. White; Christopher P. Rusconi; Bruce A. Sullenger; Frank C. Church

doi:10.1016/j.febslet.2004.04.087

RNA aptamer to thrombin binds anion-binding exosite-2 and alters protease inhibition by heparin-binding serpins

Martha L. Jeter, Linda V. Ly, Yolanda M. Fortenberry, Herbert C. Whinna, Rebekah R. White, Christopher P. Rusconi, Bruce A. Sullenger, Frank C. Church

Research output: Contribution to journal › Article › peer-review

28 Scopus citations

Abstract

We studied the RNA aptamer Toggle-25/thrombin interaction during inhibition by antithrombin (AT), heparin cofactor II (HCII) and protein C inhibitor (PCI). Thrombin inhibition was reduced 3-fold by Toggle-25 for AT and HCII, but it was slightly enhanced for PCI. In the presence of glycosaminoglycans, AT and PCI had significantly reduced thrombin inhibition with Toggle-25, but it was only reduced 3-fold for HCII. This suggested that the primary effect of aptamer binding was through the heparin-binding site of thrombin, anion-binding exosite-2 (exosite-2). We localized the Toggle-25 binding site to Arg 98, Glu 169, Lys 174, Asp 175, Arg 245, and Lys 248 of exosite-2. We conclude that a RNA aptamer to thrombin exosite-2 might provide an effective clinical reagent to control heparin's anticoagulant action.

Original language	English (US)
Pages (from-to)	10-14
Number of pages	5
Journal	FEBS Letters
Volume	568
Issue number	1-3
DOIs	https://doi.org/10.1016/j.febslet.2004.04.087
State	Published - Jun 18 2004
Externally published	Yes

Keywords

AT, antithrombin
HCII, heparin cofactor II
Heparin
PCI, protein C inhibitor
RNA aptamer
Serpin
Serpin, serine protease inhibitor
Thrombin
exosite-1 and -2, anion-binding exosite-1 and anion-binding exosite-2

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/j.febslet.2004.04.087

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@article{e0081f3a6b25432a8a9a8a20a1b33f1c,

title = "RNA aptamer to thrombin binds anion-binding exosite-2 and alters protease inhibition by heparin-binding serpins",

abstract = "We studied the RNA aptamer Toggle-25/thrombin interaction during inhibition by antithrombin (AT), heparin cofactor II (HCII) and protein C inhibitor (PCI). Thrombin inhibition was reduced 3-fold by Toggle-25 for AT and HCII, but it was slightly enhanced for PCI. In the presence of glycosaminoglycans, AT and PCI had significantly reduced thrombin inhibition with Toggle-25, but it was only reduced 3-fold for HCII. This suggested that the primary effect of aptamer binding was through the heparin-binding site of thrombin, anion-binding exosite-2 (exosite-2). We localized the Toggle-25 binding site to Arg 98, Glu 169, Lys 174, Asp 175, Arg 245, and Lys 248 of exosite-2. We conclude that a RNA aptamer to thrombin exosite-2 might provide an effective clinical reagent to control heparin's anticoagulant action.",

keywords = "AT, antithrombin, HCII, heparin cofactor II, Heparin, PCI, protein C inhibitor, RNA aptamer, Serpin, Serpin, serine protease inhibitor, Thrombin, exosite-1 and -2, anion-binding exosite-1 and anion-binding exosite-2",

author = "Jeter, {Martha L.} and Ly, {Linda V.} and Fortenberry, {Yolanda M.} and Whinna, {Herbert C.} and White, {Rebekah R.} and Rusconi, {Christopher P.} and Sullenger, {Bruce A.} and Church, {Frank C.}",

note = "Funding Information: We gratefully acknowledge Dr. Timothy Myles and Dr. Lawrence L.K. Leung (Stanford University) for providing the exosite-2 thrombin mutants used in this study. This research was supported in part by Research Grants HL-32656 and HL-06350 (to F.C.C), HL-04063 (to H.C.W.) and HL65222 (to B.A.S.) from the National Institutes of Health, and a Howard Hughes Medical Institute Postdoctoral Research Fellowship for Physicians (R.R.W). Stipend support to Y.M.F. was through the Carolina Minority Postdoctoral Scholars Program. ",

year = "2004",

month = jun,

day = "18",

doi = "10.1016/j.febslet.2004.04.087",

language = "English (US)",

volume = "568",

pages = "10--14",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "Elsevier",

number = "1-3",

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TY - JOUR

T1 - RNA aptamer to thrombin binds anion-binding exosite-2 and alters protease inhibition by heparin-binding serpins

AU - Jeter, Martha L.

AU - Ly, Linda V.

AU - Fortenberry, Yolanda M.

AU - Whinna, Herbert C.

AU - White, Rebekah R.

AU - Rusconi, Christopher P.

AU - Sullenger, Bruce A.

AU - Church, Frank C.

N1 - Funding Information: We gratefully acknowledge Dr. Timothy Myles and Dr. Lawrence L.K. Leung (Stanford University) for providing the exosite-2 thrombin mutants used in this study. This research was supported in part by Research Grants HL-32656 and HL-06350 (to F.C.C), HL-04063 (to H.C.W.) and HL65222 (to B.A.S.) from the National Institutes of Health, and a Howard Hughes Medical Institute Postdoctoral Research Fellowship for Physicians (R.R.W). Stipend support to Y.M.F. was through the Carolina Minority Postdoctoral Scholars Program.

PY - 2004/6/18

Y1 - 2004/6/18

N2 - We studied the RNA aptamer Toggle-25/thrombin interaction during inhibition by antithrombin (AT), heparin cofactor II (HCII) and protein C inhibitor (PCI). Thrombin inhibition was reduced 3-fold by Toggle-25 for AT and HCII, but it was slightly enhanced for PCI. In the presence of glycosaminoglycans, AT and PCI had significantly reduced thrombin inhibition with Toggle-25, but it was only reduced 3-fold for HCII. This suggested that the primary effect of aptamer binding was through the heparin-binding site of thrombin, anion-binding exosite-2 (exosite-2). We localized the Toggle-25 binding site to Arg 98, Glu 169, Lys 174, Asp 175, Arg 245, and Lys 248 of exosite-2. We conclude that a RNA aptamer to thrombin exosite-2 might provide an effective clinical reagent to control heparin's anticoagulant action.

AB - We studied the RNA aptamer Toggle-25/thrombin interaction during inhibition by antithrombin (AT), heparin cofactor II (HCII) and protein C inhibitor (PCI). Thrombin inhibition was reduced 3-fold by Toggle-25 for AT and HCII, but it was slightly enhanced for PCI. In the presence of glycosaminoglycans, AT and PCI had significantly reduced thrombin inhibition with Toggle-25, but it was only reduced 3-fold for HCII. This suggested that the primary effect of aptamer binding was through the heparin-binding site of thrombin, anion-binding exosite-2 (exosite-2). We localized the Toggle-25 binding site to Arg 98, Glu 169, Lys 174, Asp 175, Arg 245, and Lys 248 of exosite-2. We conclude that a RNA aptamer to thrombin exosite-2 might provide an effective clinical reagent to control heparin's anticoagulant action.

KW - AT, antithrombin

KW - HCII, heparin cofactor II

KW - Heparin

KW - PCI, protein C inhibitor

KW - RNA aptamer

KW - Serpin

KW - Serpin, serine protease inhibitor

KW - Thrombin

KW - exosite-1 and -2, anion-binding exosite-1 and anion-binding exosite-2

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U2 - 10.1016/j.febslet.2004.04.087

DO - 10.1016/j.febslet.2004.04.087

M3 - Article

C2 - 15196911

AN - SCOPUS:2942555464

SN - 0014-5793

VL - 568

SP - 10

EP - 14

JO - FEBS Letters

JF - FEBS Letters

IS - 1-3

ER -

RNA aptamer to thrombin binds anion-binding exosite-2 and alters protease inhibition by heparin-binding serpins

Abstract

Keywords

ASJC Scopus subject areas

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