title: Molecular genetic analysis of a variant Bernard-Soulier syndrome due to compound heterozygosity for two novel glycoprotein Ibbeta mutations.
abstract: Bernard-Soulier syndrome (BSS) is a rare bleeding disorder characterized by giant platelets, thrombocytopenia, and prolonged bleeding time. It is caused by abnormalities in the glycoprotein (GP) Ib/IX/V complex, the receptor for von Willebrand factor (vWF). Most cases of BSS described so far involve quantitative rather than qualitative defects in the complex. In this study, we investigated the effects of two naturally occurring mutations in the GPIbbeta gene, C122S and 443delG, on the expression of the GPIb/IX complex identified in a variant type of BSS in which the platelets had severely reduced GPIbalpha ( approximately 10%) and less markedly reduced GPIbbeta and GPIX ( approximately 20%) expression. Immunoblot analysis showed the absence of non-reduced GPIb (GPIbalpha/GPIbbeta) in the patient's platelets. Transient transfection experiments in 293T cells revealed the expression of GPIbbeta Ser122 polypeptide and absence of GPIbbeta 443delG polypeptide. Although no disulfide-linked association was observed between GPIbbeta Ser122 and GPIbalpha, GPIbbeta Ser122 was non-covalently associated with both GPIbalpha and GPIX subunits on the cell surface when cotransfected with wild-type GPIbalpha and GPIX. Chinese hamster ovary cells stably expressing GPIbalpha/Ibbeta Ser122/IX had the ability to bind soluble vWF and to aggregate in the presence of ristocetin. These results suggest that despite disruption of the disulfide linkage between GPIbalpha and GPIbbeta, GPIb/IX is formed, but its stability may be impaired, resulting in low levels of the complex on the platelet membranes.
authors.affiliation: Department of Hemostasis and Thrombosis, Clinical Research Center, National Hospital Organization Nagoya Medical Center, Nagoya, Japan. kunishis@nnh.hosp.go.jp
authors.forename: Shinji
authors.initials: S
authors.lastname: Kunishima
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sako
authors.forename: Tomio
authors.initials: T
authors.lastname: Yamazaki
authors.forename: Motohiro
authors.initials: M
authors.lastname: Hamaguchi
authors.forename: Hidehiko
authors.initials: H
authors.lastname: Saito
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001606:Bernard-Soulier Syndrome / Q000235:genetics*; D001792:Blood Platelets / Q000378:metabolism; D016466:CHO Cells; D002648:Child; D006224:Cricetinae; D006579:Heterozygote; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000235:genetics*; D013921:Thrombocytopenia / Q000235:genetics
chemical_list: D019038:Platelet Glycoprotein GPIb-IX Complex