title: HRG Tokushima: molecular and cellular characterization of histidine-rich glycoprotein (HRG) deficiency.
abstract: Previously, we found the first congenital deficiency of histidine-rich glycoprotein (HRG) in a Japanese woman with thrombosis. To elucidate the genetic basis of this deficiency, we first performed Southern blot analysis and found no gross deletion or insertion in the proband's HRG gene. We then examined the nucleotide sequences of all seven exons of the proband's HRG gene. A single nucleotide substitution, G to A at nucleotide position 429, which mutates Gly85 to Glu in the first cystatin-like domain, was found in exon 3 in 13 of 22 amplified clones. This mutation generates a unique Taq I site. Exon 3 was amplified from the proband, her family members, and 50 unrelated normal Japanese individuals, and Taq I fragmentation was examined. Fragmentation of exon 3 was observed in one allele of the genes from the proband and the family members who also have decreased plasma levels of HRG. Fifty unrelated normal Japanese individuals had a normal HRG gene, indicating that the G to A mutation is not a common polymorphism. To elucidate the identified mutation as a cause for the secretion defect of HRG in the proband's plasma, we constructed and transiently expressed the recombinant Tokushima-type HRG mutant (Gly85 to Glu) in baby hamster kidney (BHK) cells, and examined an intracellular event of the mutant protein. The results showed that only about 20% of the Tokushima-type HRG was secreted into the culture medium, and intracellular degradation of the mutant was observed. Thus, the present study strongly suggests that the HRG deficiency is caused by intracellular degradation of the Gly85 to Glu mutant of HRG in the proband.
authors.affiliation: First Department of Internal Medicine, School of Medicine, The University of Tokushima, Tokushima, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Shigekiyo
authors.forename: H
authors.initials: H
authors.lastname: Yoshida
authors.forename: K
authors.initials: K
authors.lastname: Matsumoto
authors.forename: H
authors.initials: H
authors.lastname: Azuma
authors.forename: S
authors.initials: S
authors.lastname: Wakabayashi
authors.forename: S
authors.initials: S
authors.lastname: Saito
authors.forename: K
authors.initials: K
authors.lastname: Fujikawa
authors.forename: T
authors.initials: T
authors.lastname: Koide
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D003241:Consanguinity; D003270:Contraceptive Agents / Q000009:adverse effects; D006224:Cricetinae; D005260:Female; D006801:Humans; D008647:Mesocricetus; D008969:Molecular Sequence Data; D010375:Pedigree; D011506:Proteins / Q000235:genetics*; D011817:Rabbits; D051381:Rats; D012851:Sinus Thrombosis, Intracranial / Q000139:chemically induced / Q000235:genetics; D019851:Thrombophilia / Q000150:complications / Q000235:genetics*; D000099053:Histidine-Rich Glycoprotein
chemical_list: D003270:Contraceptive Agents; D011506:Proteins; D000099053:Histidine-Rich Glycoprotein