Molecular bases of defective signal transduction in the platelet P2Y12 receptor of a patient with congenital bleeding

Marco Cattaneo, Maddalena L. Zighetti, Rossana Lombardi, Constantino Martinez, Anna Lecchi, Pamela B. Conley, Jerry Ware, Zaverio M. Ruggeri

Research output: Contribution to journalArticle

152 Citations (Scopus)

Abstract

We have identified structural attributes required for signal transduction through a seven-transmembrane-domain receptor. Platelets from a patient (AC) with a congenital bleeding disorder had normal shape change but reduced and reversible aggregation in response to 4 μM ADP, similar to normal platelets with blocked P2Y12 receptor. The response to 20 μM ADP, albeit still decreased, was more pronounced and was reduced by a P2Y12 antagonist, indicating some residual receptor function. ADP failed to lower the adenylyl cyclase activity stimulated by prostaglandin E1 in the patient's platelets, even though the number and affinity of 2-methylthioadenosine 5′-[33P]diphosphate-binding sites was normal. Analysis of the patient's P2Y12 gene revealed a G-to-A transition in one allele, changing the codon for Arg-256 in the sixth transmembrane domain to Gln, and a C-to-T transition in the other allele, changing the codon for Arg-265 in the third extracellular loop to Trp. Neither mutation interfered with receptor surface expression but both altered function, since ADP inhibited the forskolin-induced increase of cAMP markedly less in cells transfected with either mutant P2Y12 as compared with wild-type receptor. These studies delineate a region of P2Y12 required for normal function after ADP binding.

Original languageEnglish
Pages (from-to)1978-1983
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume100
Issue number4
DOIs
Publication statusPublished - Feb 18 2003

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Adenosine Diphosphate
Signal Transduction
Blood Platelets
Hemorrhage
Codon
Alleles
Congenital, Hereditary, and Neonatal Diseases and Abnormalities
Diphosphates
Alprostadil
Colforsin
Platelet Count
Adenylyl Cyclases
Binding Sites
Mutation
Genes

Keywords

  • ADP
  • G-protein coupled receptors
  • Platelet aggregation
  • Platelet function disorder

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Molecular bases of defective signal transduction in the platelet P2Y12 receptor of a patient with congenital bleeding. / Cattaneo, Marco; Zighetti, Maddalena L.; Lombardi, Rossana; Martinez, Constantino; Lecchi, Anna; Conley, Pamela B.; Ware, Jerry; Ruggeri, Zaverio M.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, No. 4, 18.02.2003, p. 1978-1983.

Research output: Contribution to journalArticle

Cattaneo, Marco ; Zighetti, Maddalena L. ; Lombardi, Rossana ; Martinez, Constantino ; Lecchi, Anna ; Conley, Pamela B. ; Ware, Jerry ; Ruggeri, Zaverio M. / Molecular bases of defective signal transduction in the platelet P2Y12 receptor of a patient with congenital bleeding. In: Proceedings of the National Academy of Sciences of the United States of America. 2003 ; Vol. 100, No. 4. pp. 1978-1983.
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