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PMID: 8280110 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Different pathways for control of Na+/H+ exchange via activation of the thrombin receptor.

The Biochemical journal ·Vol. 297 ( Pt 1) ·1994-01-01 ·Pages 47-52

Nieuwland R, van Willigen G, Akkerman JW

Abstract

The aim of the present study was to clarify the control of Na+/H+ exchange in platelets activated via the thrombin receptor. When human BCECF-loaded platelets were stimulated with the thrombin-receptor-activating peptide (TRAP; amino acid sequence SFLLRN), which activates the receptor independently of proteolysis, the cytosolic pH (pHi) rose from 7.13 +/- 0.04 (n = 6) to 7.27 +/- 0.04 (n = 5), followed by a rapid decrease to resting values. Trypsin, which cleaves the receptor, induced a rapid and irreversible rise in pHi to 7.31 +/- 0.06 (n = 5). gamma-Thrombin, which cleaves the receptor but is unable to bind to the hirudin-like domain, induced a slow and irreversible rise in pHi to 7.31 +/- 0.04 (n = 14). alpha-Thrombin, which cleaves the receptor and binds to its hirudin-like domain, induced a rapid and irreversible rise in pHi to 7.31 +/- 0.04 (n = 22). Changes in pHi induced by TRAP, trypsin, gamma- and alpha-thrombin were accompanied by similar changes in cytosolic Ca2+ concentration ([Ca2+]i) and 32P-pleckstrin, a substrate of protein kinase C (PKC). The separate chelation of Ca2+i (30 microM BAPTA-AM) or inhibition of PKC (1 microM staurosporine) induced about 50% inhibition of the pHi responses triggered by TRAP, trypsin, gamma- and alpha-thrombin, but the combination induced complete inhibition. Thus the different types of activation of the thrombin receptor control Na+/H+ exchange via the same mechanism. Binding of thrombin to the hirudin-like domain accelerates exchange activation, whereas proteolysis of the receptor is essential for a sustained increase in pHi.

MeSH Terms
Alkaloids/pharmacology Amino Acid Sequence Blood Platelets/drug effects,metabolism Blood Proteins/metabolism Calcium/blood Egtazic Acid/analogs & derivatives,pharmacology Fluoresceins/pharmacology Humans Hydrogen-Ion Concentration Molecular Sequence Data Peptide Fragments/pharmacology Phosphoproteins Protein Kinase C/antagonists & inhibitors Receptors, Thrombin/drug effects,physiology Sodium-Hydrogen Exchangers/blood Staurosporine Thrombin/pharmacology Trypsin/pharmacology
Chemicals
Alkaloids Blood Proteins Fluoresceins Peptide Fragments Phosphoproteins Receptors, Thrombin Sodium-Hydrogen Exchangers platelet protein P47 thrombin receptor peptide (42-55) Egtazic Acid 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein Protein Kinase C Trypsin Thrombin Staurosporine 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid Calcium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Nieuwland R
Department of Haematology, University Hospital Utrecht, The Netherlands.
van Willigen G
Akkerman J W
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2970047
Published
1994-01-01
Pages
47-52
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1137788
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2970047 PMC Full Text View journal details
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