Molecular mechanisms of platelet exocytosis: Requirements for α-granule release

Paula P. Lemons; Dong Chen; Sidney W. Whiteheart

doi:10.1006/bbrc.1999.2039

Molecular mechanisms of platelet exocytosis: Requirements for α-granule release

Paula P. Lemons, Dong Chen, Sidney W. Whiteheart

Molecular and Cellular Biochemistry

Producción científica: Article › revisión exhaustiva

94 Citas (Scopus)

Resumen

Platelets function by secreting components necessary for primary clot formation. This report describes an in vitro assay that measures α-granule secretion. Using permeabilized platelets, it is possible to recreate Ca²⁺-stimulated release of platelet factor 4 (PF4) that is ATP- and temperature-dependent. Though other divalent cations can replace Ca²⁺ (i.e., Sr²⁺, Mn²⁺, Zn²⁺), there is no effect of Ba²⁺. Analysis by electron microscopy indicates that the in vitro assay also mimics the cytoskeletal rearrangements and granule centralization that occurs upon platelet activation in vivo. Antibody inhibition studies show that PF4 release requires the general membrane fusion protein N-ethylmaleimide-sensitive factor (NSF) and well as the target membrane SNAP receptors (t-SNAREs), syntaxin 2, 4, and SNAP-23. As shown by electron microscopy, the anti-t-SNARE antibodies block granule to target membrane fusion. This finding is unique in that it is the first report of a role for two syntaxins in the same exocytosis event. (C) 2000 Academic Press.

Idioma original	English
Páginas (desde-hasta)	875-880
Número de páginas	6
Publicación	Biochemical and Biophysical Research Communications
Volumen	267
N.º	3
DOI	https://doi.org/10.1006/bbrc.1999.2039
Estado	Published - ene 27 2000

Nota bibliográfica

Funding Information:
The authors thank the staff of the Central Kentucky Blood Center for their assistance and the members of the Whiteheart lab for their helpful discussions. We especially thank Dr. Bruce Maley, Mary Gail Engle, and Richard Watson at the College of Medicine Imaging Facility, University of Kentucky, for their assistance with the electron microscopy studies. This work was supported by Grant HL56652 from the Heart, Lung, and Blood Institute of the National Institutes of Health.

Financiación

The authors thank the staff of the Central Kentucky Blood Center for their assistance and the members of the Whiteheart lab for their helpful discussions. We especially thank Dr. Bruce Maley, Mary Gail Engle, and Richard Watson at the College of Medicine Imaging Facility, University of Kentucky, for their assistance with the electron microscopy studies. This work was supported by Grant HL56652 from the Heart, Lung, and Blood Institute of the National Institutes of Health.

Financiadores	Número del financiador
National Heart, Lung, and Blood Institute (NHLBI)	R01HL056652

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

Acceder al documento

10.1006/bbrc.1999.2039

Otros archivos y enlaces

Citar esto

@article{36b84790e44e421fb280d1f73b73c280,

title = "Molecular mechanisms of platelet exocytosis: Requirements for α-granule release",

abstract = "Platelets function by secreting components necessary for primary clot formation. This report describes an in vitro assay that measures α-granule secretion. Using permeabilized platelets, it is possible to recreate Ca2+-stimulated release of platelet factor 4 (PF4) that is ATP- and temperature-dependent. Though other divalent cations can replace Ca2+ (i.e., Sr2+, Mn2+, Zn2+), there is no effect of Ba2+. Analysis by electron microscopy indicates that the in vitro assay also mimics the cytoskeletal rearrangements and granule centralization that occurs upon platelet activation in vivo. Antibody inhibition studies show that PF4 release requires the general membrane fusion protein N-ethylmaleimide-sensitive factor (NSF) and well as the target membrane SNAP receptors (t-SNAREs), syntaxin 2, 4, and SNAP-23. As shown by electron microscopy, the anti-t-SNARE antibodies block granule to target membrane fusion. This finding is unique in that it is the first report of a role for two syntaxins in the same exocytosis event. (C) 2000 Academic Press.",

keywords = "NSF, Platelets, SNAREs, Syntaxin, α-granules",

author = "Lemons, \{Paula P.\} and Dong Chen and Whiteheart, \{Sidney W.\}",

note = "Funding Information: The authors thank the staff of the Central Kentucky Blood Center for their assistance and the members of the Whiteheart lab for their helpful discussions. We especially thank Dr. Bruce Maley, Mary Gail Engle, and Richard Watson at the College of Medicine Imaging Facility, University of Kentucky, for their assistance with the electron microscopy studies. This work was supported by Grant HL56652 from the Heart, Lung, and Blood Institute of the National Institutes of Health.",

year = "2000",

month = jan,

day = "27",

doi = "10.1006/bbrc.1999.2039",

language = "English",

volume = "267",

pages = "875--880",

number = "3",

}

TY - JOUR

T1 - Molecular mechanisms of platelet exocytosis

T2 - Requirements for α-granule release

AU - Lemons, Paula P.

AU - Chen, Dong

AU - Whiteheart, Sidney W.

N1 - Funding Information: The authors thank the staff of the Central Kentucky Blood Center for their assistance and the members of the Whiteheart lab for their helpful discussions. We especially thank Dr. Bruce Maley, Mary Gail Engle, and Richard Watson at the College of Medicine Imaging Facility, University of Kentucky, for their assistance with the electron microscopy studies. This work was supported by Grant HL56652 from the Heart, Lung, and Blood Institute of the National Institutes of Health.

PY - 2000/1/27

Y1 - 2000/1/27

N2 - Platelets function by secreting components necessary for primary clot formation. This report describes an in vitro assay that measures α-granule secretion. Using permeabilized platelets, it is possible to recreate Ca2+-stimulated release of platelet factor 4 (PF4) that is ATP- and temperature-dependent. Though other divalent cations can replace Ca2+ (i.e., Sr2+, Mn2+, Zn2+), there is no effect of Ba2+. Analysis by electron microscopy indicates that the in vitro assay also mimics the cytoskeletal rearrangements and granule centralization that occurs upon platelet activation in vivo. Antibody inhibition studies show that PF4 release requires the general membrane fusion protein N-ethylmaleimide-sensitive factor (NSF) and well as the target membrane SNAP receptors (t-SNAREs), syntaxin 2, 4, and SNAP-23. As shown by electron microscopy, the anti-t-SNARE antibodies block granule to target membrane fusion. This finding is unique in that it is the first report of a role for two syntaxins in the same exocytosis event. (C) 2000 Academic Press.

AB - Platelets function by secreting components necessary for primary clot formation. This report describes an in vitro assay that measures α-granule secretion. Using permeabilized platelets, it is possible to recreate Ca2+-stimulated release of platelet factor 4 (PF4) that is ATP- and temperature-dependent. Though other divalent cations can replace Ca2+ (i.e., Sr2+, Mn2+, Zn2+), there is no effect of Ba2+. Analysis by electron microscopy indicates that the in vitro assay also mimics the cytoskeletal rearrangements and granule centralization that occurs upon platelet activation in vivo. Antibody inhibition studies show that PF4 release requires the general membrane fusion protein N-ethylmaleimide-sensitive factor (NSF) and well as the target membrane SNAP receptors (t-SNAREs), syntaxin 2, 4, and SNAP-23. As shown by electron microscopy, the anti-t-SNARE antibodies block granule to target membrane fusion. This finding is unique in that it is the first report of a role for two syntaxins in the same exocytosis event. (C) 2000 Academic Press.

KW - NSF

KW - Platelets

KW - SNAREs

KW - Syntaxin

KW - α-granules

UR - https://www.scopus.com/pages/publications/0034719338

UR - https://www.scopus.com/inward/citedby.url?scp=0034719338&partnerID=8YFLogxK

U2 - 10.1006/bbrc.1999.2039

DO - 10.1006/bbrc.1999.2039

M3 - Article

C2 - 10673384

AN - SCOPUS:0034719338

SN - 0006-291X

VL - 267

SP - 875

EP - 880

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 3

ER -

Molecular mechanisms of platelet exocytosis: Requirements for α-granule release

Resumen

Nota bibliográfica

Financiación

ASJC Scopus subject areas

Acceder al documento

Otros archivos y enlaces

Huella

Citar esto