Faculty Collaboration Database

Cloning and expression of canine glycoprotein Ibalpha. Thromb Haemost 1999 Oct;82(4):1327-33

Date

11/02/1999

Pubmed ID

10544923

Scopus ID

2-s2.0-0032827491 (requires institutional sign-in at Scopus site)   8 Citations

Abstract

The interaction of the glycoprotein (GP) Ib-IX-V complex with von Willebrand factor (vWF) is critical in initiation of haemostasis and thrombosis through platelet adhesion to damaged endothelium. The binding site for vWF resides within the GPIbalpha subunit of the complex. To further define the physiological function of platelet GPIbalpha we cloned and expressed the canine GPIbalpha cDNA. A canine platelet cDNA library was constructed and screened with a randomly primed 32P-labeled 1041-base-pair restriction fragment of the human GPIbalpha cDNA. Analysis of 23 clones demonstrated that the canine GPIbalpha cDNA is 2530 nucleotides in length and includes a short 5' untranslated segment of 42 nucleotides followed by a signal peptide of 16 amino acids, a mature peptide of 645 amino acids and a 3' noncoding region of 455 nucleotides. A single intron of 142 nucleotides, 6 nucleotides upstream from the ATG translation initiation codon was identified in the canine gene in a similar location to that present in the human gene. Chinese hamster ovary cells that stably express human GPIbbeta and GPIX were transfected with the canine GPIbalpha cDNA. Canine GPIbalpha was expressed on the surface of these cells and bound vWF in the presence of botrocetin. The binding of vWF was inhibited by an anti-vWF human monoclonal antibody known to inhibit vWF binding to GPIbalpha. The results of this investigation will allow the development of reagents to study the physiological function of GPIbalpha in an animal model.

Author List

Kenny D, Morateck PA, Fahs SA, Warltier DC, Montgomery RR

MESH terms used to index this publication - Major topics in bold

Amino Acid Sequence
Animals
Base Sequence
Cloning, Molecular
Cricetinae
DNA, Complementary
Dogs
Humans
Molecular Sequence Data
Platelet Glycoprotein GPIb-IX Complex
Sequence Alignment