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The human platelet alloantigens, HPA‐5(a+ b‐) and HPA‐5(a‐ b+), are associated with a Glu 505 /Lys 505 polymorphism of glycoprotein Ia (the α 2 subunit of VLA‐2)
Author(s) -
Simsek S.,
Gallardo D.,
Ribera A.,
Borne A. E. G. Kr.
Publication year - 1994
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1994.tb04808.x
Subject(s) - platelet membrane glycoprotein , complementary dna , microbiology and biotechnology , biology , polymerase chain reaction , glycoprotein , platelet , allele , genetics , gene , immunology
Summary. GP Ia/IIa (also called VLA‐2 or α2β1) is the primary receptor for collagen on platelets. The human platelet alloantigens HPA‐5a(Br b ) and HPA‐5b(Br a ) have been found to reside on the platelet GP Ia/IIa complex. In order to establish the molecular basis of the HPA‐5 system, platelet RNA was isolated from HPA‐5 (a+ b‐) and HPA‐5(a b+) individuals. After reverse transcription, cDNA coding for glycoprotein Ia (GP Ia) was amplified by the polymerase chain reaction (PCR). Nucleotide sequence analysis of the PCR products revealed an A·G polymorphism at base pair 1648 of the coding region of the mature protein, resulting in a substitution of lysine (AAG) in HPA‐5b(Br a ) by glutamic acid (GAG) in HPA‐5a (Br b ) at amino acid 505. Subsequent PCR‐ASRA (allele‐specific restriction enzyme analysis) with Mnl I using cDNA derived from three HPA‐5 (a+b‐), one HPA‐5 (a+b+) individuals demonstrated that HPA‐5a and −5b alleles are distinguishable by DNA typing. In addition to the A→G substitution at base pair 1648, three silent mutations were identified, G→C (195 bp), C→T (837 bp), G→A (1041 bp).