Abstract
The platelet glycoprotein (GP) Ib-IX-V complex mediates adhesion to von Willebrand factor (vWf) in (patho)physiologic thrombus formation. The vWf- binding site on GP Ib-IX-V is within the N-terminal 282 residues of GP Ibα, which consist of an N-terminal flanking sequence (His-1-IIe-35), 7 leucine- rich repeats (Leu-36-Ala200), a C-terminal flank (Phe-201-Gly268), and a sulfated tyrosine sequence (Asp-269-Glu-282). We have used mammalian cell expression of canine-human chimeras of GP Ibα, corresponding to precise structural boundaries, to demonstrate the first specific requirement for individual leucine-rich repeats for binding of vWf either induced by a modulator, ristocetin, or under hydrodynamic flow. Implicit in this approach was that the GP Ibα chimeras retained a functional conformation, a supposition confirmed by analyzing restoration of function to reversed human- canine chimeras and demonstrating that all chimeras bound vWf activated by botrocetin, a modulator that is indiscriminate between species. Leucine-rich repeats 2, 3, and 4 of GP Ibα were identified as being critical for vWf adhesion to GP Ib-IX-V.
| Original language | English |
|---|---|
| Pages (from-to) | 903-910 |
| Number of pages | 8 |
| Journal | Blood |
| Volume | 95 |
| Issue number | 3 |
| Publication status | Published - 1 Feb 2000 |
Cite this
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Requirement of leucine-rich repeats of glycoprotein (GP) Ibα for shear- dependent and static binding of von willebrand factor to the platelet membrane GP Ib-IX-V complex. / Shen, Yang; Romo, Gabriel M.; Dong, Jing Fei; Schade, Alicia; McIntire, Larry V.; Kenny, Dermot; Whisstock, James C.; Berndt, Michael C.; López, José A.; Andrews, Robert K.
In: Blood, Vol. 95, No. 3, 01.02.2000, p. 903-910.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Requirement of leucine-rich repeats of glycoprotein (GP) Ibα for shear- dependent and static binding of von willebrand factor to the platelet membrane GP Ib-IX-V complex
AU - Shen, Yang
AU - Romo, Gabriel M.
AU - Dong, Jing Fei
AU - Schade, Alicia
AU - McIntire, Larry V.
AU - Kenny, Dermot
AU - Whisstock, James C.
AU - Berndt, Michael C.
AU - López, José A.
AU - Andrews, Robert K.
PY - 2000/2/1
Y1 - 2000/2/1
N2 - The platelet glycoprotein (GP) Ib-IX-V complex mediates adhesion to von Willebrand factor (vWf) in (patho)physiologic thrombus formation. The vWf- binding site on GP Ib-IX-V is within the N-terminal 282 residues of GP Ibα, which consist of an N-terminal flanking sequence (His-1-IIe-35), 7 leucine- rich repeats (Leu-36-Ala200), a C-terminal flank (Phe-201-Gly268), and a sulfated tyrosine sequence (Asp-269-Glu-282). We have used mammalian cell expression of canine-human chimeras of GP Ibα, corresponding to precise structural boundaries, to demonstrate the first specific requirement for individual leucine-rich repeats for binding of vWf either induced by a modulator, ristocetin, or under hydrodynamic flow. Implicit in this approach was that the GP Ibα chimeras retained a functional conformation, a supposition confirmed by analyzing restoration of function to reversed human- canine chimeras and demonstrating that all chimeras bound vWf activated by botrocetin, a modulator that is indiscriminate between species. Leucine-rich repeats 2, 3, and 4 of GP Ibα were identified as being critical for vWf adhesion to GP Ib-IX-V.
AB - The platelet glycoprotein (GP) Ib-IX-V complex mediates adhesion to von Willebrand factor (vWf) in (patho)physiologic thrombus formation. The vWf- binding site on GP Ib-IX-V is within the N-terminal 282 residues of GP Ibα, which consist of an N-terminal flanking sequence (His-1-IIe-35), 7 leucine- rich repeats (Leu-36-Ala200), a C-terminal flank (Phe-201-Gly268), and a sulfated tyrosine sequence (Asp-269-Glu-282). We have used mammalian cell expression of canine-human chimeras of GP Ibα, corresponding to precise structural boundaries, to demonstrate the first specific requirement for individual leucine-rich repeats for binding of vWf either induced by a modulator, ristocetin, or under hydrodynamic flow. Implicit in this approach was that the GP Ibα chimeras retained a functional conformation, a supposition confirmed by analyzing restoration of function to reversed human- canine chimeras and demonstrating that all chimeras bound vWf activated by botrocetin, a modulator that is indiscriminate between species. Leucine-rich repeats 2, 3, and 4 of GP Ibα were identified as being critical for vWf adhesion to GP Ib-IX-V.
UR - http://www.scopus.com/inward/record.url?scp=12944317287&partnerID=8YFLogxK
M3 - Article
VL - 95
SP - 903
EP - 910
JO - Blood
JF - Blood
SN - 0006-4971
IS - 3
ER -