title: Soluble expression of human glycoprotein Ibα in Escherichia coli through replacement of the N-terminal capping domain.
abstract: Glycoprotein Ibα (GpIbα), a family of LRR (leucine-rich repeat) proteins, is a membrane protein on the platelet, and plays an important role in atherothrombotic events. The complex formation of GpIbα with the von Willebrand Factor (vWF) has been revealed to lead to acute coronary syndrome (ACS) or stroke. A considerable attention has been paid to understand the biological functions of GpIbα and its regulation. However, difficulty with the soluble expression of human GpIbα in bacteria has hampered the relevant research. Herein, we present a soluble expression of GpIbα in Escherichiacoli by replacing the N-terminal capping domain of GpIbα with that of Internalin B using a computational approach. The resulting protein was expressed as a soluble form in E. coli, maintaining its structural feature and binding property for vWF. The present approach can be broadly used for the soluble expression of human LRR proteins in E. coli.
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 373-1, Guseong-Dong, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Jeong-Hyun
authors.initials: JH
authors.lastname: Ryou
authors.affiliation: Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), 373-1, Guseong-Dong, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Keunwan
authors.initials: K
authors.lastname: Park
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 373-1, Guseong-Dong, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Joong-Jae
authors.initials: JJ
authors.lastname: Lee
authors.affiliation: Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), 373-1, Guseong-Dong, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Dongsup
authors.initials: D
authors.lastname: Kim
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), 373-1, Guseong-Dong, Yuseong-gu, Daejeon 305-701, Republic of Korea. Electronic address: hskim76@kaist.ac.kr.
authors.forename: Hak-Sung
authors.initials: HS
authors.lastname: Kim
mesh_terms: D000906:Antibodies / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D014841:von Willebrand Factor / Q000737:chemistry*
chemical_list: D000906:Antibodies; D001426:Bacterial Proteins; D008565:Membrane Proteins; D019038:Platelet Glycoprotein GPIb-IX Complex; D011993:Recombinant Fusion Proteins; C094990:adhesion receptor; C095768:inlB protein, Listeria monocytogenes; D014841:von Willebrand Factor
keywords: Glycoprotein Ibα; Internalin B; N-terminal capping domain; Repeat protein