T-shaped arrangement of the recombinant agrin G3 - IgG Fc protein

Trushar R. Patel, Markus Meier, Jianhua Li, Gordon Morris, Arthur J. Rowe, Jörg Stetefeld

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Agrin is a large heparin sulphate proteoglycan with multiple domains, which is located in the extracellular matrix. The C-terminal G3 domain of agrin is functionally one of the most important domains. It harbors an α-dystroglycan binding site and carries out acetylcholine receptor clustering activities. In the present study, we have fused the G3 domain of agrin to an IgG Fc domain to produce a G3-Fc fusion protein that we intend to use as a tool to investigate new binding partners of agrin. As a first step of the study, we have characterized the recombinant fusion protein using a multidisciplinary approach using dynamic light scattering, analytical ultracentrifugation and small angle X-ray scattering (SAXS). Interestingly, our SAXS analysis using the high-resolution structures of G3 and Fc domain as models indicates that the G3-Fc protein forms a T-shaped molecule with the G3 domains extruding perpendicularly from the Fc scaffold. To validate our models, we have used the program HYDROPRO to calculate the hydrodynamic properties of the solution models. The calculated values are in excellent agreement with those determined experimentally.

Original languageEnglish
Pages (from-to)931-940
Number of pages10
JournalProtein Science
Volume20
Issue number6
Early online date26 Apr 2011
DOIs
Publication statusPublished - Jun 2011

Fingerprint

Agrin
Immunoglobulin G
X ray scattering
Proteins
X-Rays
Dystroglycans
Recombinant Fusion Proteins
Ultracentrifugation
Cholinergic Receptors
Dynamic light scattering
Hydrodynamics
Ports and harbors
Scaffolds
Extracellular Matrix
Cluster Analysis
Fusion reactions
Binding Sites
Molecules

Cite this

Patel, T. R., Meier, M., Li, J., Morris, G., Rowe, A. J., & Stetefeld, J. (2011). T-shaped arrangement of the recombinant agrin G3 - IgG Fc protein. Protein Science, 20(6), 931-940. https://doi.org/10.1002/pro.628
Patel, Trushar R. ; Meier, Markus ; Li, Jianhua ; Morris, Gordon ; Rowe, Arthur J. ; Stetefeld, Jörg. / T-shaped arrangement of the recombinant agrin G3 - IgG Fc protein. In: Protein Science. 2011 ; Vol. 20, No. 6. pp. 931-940.
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Patel, TR, Meier, M, Li, J, Morris, G, Rowe, AJ & Stetefeld, J 2011, 'T-shaped arrangement of the recombinant agrin G3 - IgG Fc protein', Protein Science, vol. 20, no. 6, pp. 931-940. https://doi.org/10.1002/pro.628

T-shaped arrangement of the recombinant agrin G3 - IgG Fc protein. / Patel, Trushar R.; Meier, Markus; Li, Jianhua; Morris, Gordon; Rowe, Arthur J.; Stetefeld, Jörg.

In: Protein Science, Vol. 20, No. 6, 06.2011, p. 931-940.

Research output: Contribution to journalArticle

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