Print Email Facebook Twitter Identification and genetic analysis of a common molecular variant of histidine-rich glycoprotein with a difference of 2KD in apparent molecular weight Title Identification and genetic analysis of a common molecular variant of histidine-rich glycoprotein with a difference of 2KD in apparent molecular weight Author Hennis, B.C. van Boheemen, P.A. Wakabayashi, S. Koide, T. Hoffmann, J.J.M.L. Kievit, P. Dooijewaard, G. Jansen, J.G. Klutf, C. TNO Preventie en Gezondheid Publication year 1995 Abstract Two forms of histidine-rich glycoprotein (HRG) were detected on SDS-PAGE by silver staining and immunoblotting after isolation of the protein from pooled plasma using immune-affinity chromatography followed by chromatography with heparin-Sepharose. Both forms were single-chain molecules and the apparent molecular weights of form 1 and form 2 were 77 kD and 75 kD respectively. Mendelian inheritance of both HRG forms was observed in four families with 24 informative meioses, strongly suggesting that the two forms are encoded by different alleles. The frequency of form 1 and form 2 in a group of 36 individuals was 0.35 and 0.65 respectively. The difference between the two molecular variants was studied by direct sequence analysis of amplified exons of the HRG gene from 6 individuals who were homozygous either for form 1 or form 2. Five amino acid polymorphisms in three different exons were observed: Ile/Thr in exon 4; Pro/Ser in exon 5; His/Arg, Arg/Cys and Asn/lle in exon 7. Analysis of these polymorphisms in 20 volunteers showed that only the Pro/Ser polymorphism at position 186 in exon 5 was coupled to the form of the HRG protein. Ser was found in form 1 and Pro in form 2. The presence of Ser at position 186 introduces a consensus sequence for a N-glycosylation site (Asn-X-Ser/Thr). By removing N-linked sugars with N-glycanase, it could be demonstrated that the difference between the two forms of HRG is caused by an extra carbohydrate group at Asn 184 in form 1. Subject Biologyglycoproteinhistidineprolineserineallelearticleclinical articleexonfamily studygene amplificationgene frequencygenetic analysisgenetic polymorphismhomozygosityhumanimmunoblottinginheritancemolecular weightpolyacrylamide gel electrophoresispriority journalprotein variantsequence analysissilver stainingAmino Acid SequenceBase SequenceBlood ProteinsCase-Control StudiesExonsGenetic CodeGenotypeGlycoproteinsHumansMolecular Sequence DataMolecular WeightPedigreePhenotypePolymorphism, GeneticProteinsVariation (Genetics) To reference this document use: http://resolver.tudelft.nl/uuid:0bb393f2-d1df-4dcd-a1eb-17ae4896a247 TNO identifier 233104 ISSN 0340-6245 Source Thrombosis and Haemostasis, 74 (6), 1491-1496 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.