Histidine-tagging effect on TGF-β molecular dynamics

Authors

1 Tarbiat Modares University

2 Tarbiat Modares university

3 Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

Abstract
Transforming growth factor beta (TGF-β), is a small homodimeric signaling protein. The TGF-β isoforms (TGFβ1, β2 and β3) are involved in many cellular processes including growth inhibition, extracellular matrix remodeling, tissue development, cell migration, invasion and immune regulation. For research aims, TGFβs are overexpressed using recombinant eukaryotic cell or bacterial expression systems. For achieving an efficient purification of TGF-β by immobilized metal ion affinity chromatography (IMAC), a histidine tag was placed either at the C-terminal (C-TGFβ) or N-terminal (N-TGFβ) region of the sequence and the effect of His-tag on TGF-β structure has been studied by computational tools. Proteins 3D structures were modeled using MODELLER software and molecular dynamics simulation of native TGF-β and modelled proteins, N-TGFβ and C-TGFβ were studied in water by GROMACS package. Protein dynamics modeling indicated that the His-tag attached at the C-terminus but not at the N-terminus of the TGF-β can affect the fluctuations of amino acids and protein structure. It is concluded that the C-terminal tagging may cause distortion and misfolding in the structure.

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[1] Massagué, J. (1998) TGF-β signal transduction,
Annual Review of Biochemistry, USA, pp 753-791.
[2] Derynck, R., Zhang, Y. E. (2003) Smaddependent and Smad-independent pathways in
TGF-beta family signalling. Nature. 425. 577-584.
[3] Daopin, S., Piez, K. A., Ogawa, Y., Davies ,D.
R. (1992) Crystal structure of transforming growth
factor-b2: an unusual fold for the superfamily,
Science. 257. 369-373.
[4] Hinck, A. P., Archer, S. J., Qian, S. W.,
Roberts, A. B., Sporn, M. B., Weatherbee, J. A.,
Tsang, M. L. S., Lucas, R., Zhang, B. L., Wenker,
J. (1996) Transforming growth factor β1: threedimensional structure in solution and comparison
with the X-ray structure of transforming growth
factor β2, Biochemistry. 35, 8517-8534.
[5] Mittl, P. R., Priestle, J. P., Cox, D. A.,
Mcmaster, G., Cerletti, N., Grütter, M. G. (1996)
The crystal structure of TGF‐β3 and comparison to
TGF‐β2: implications for receptor binding, Protein
Sci. 5, 1261-1271.
[6] Radaev, S., Zou, Z., Huang, T., Lafer, E. M.,
Hinck, A. P., Sun, P. D. (2010) Ternary complex of
transforming growth factor-β1 reveals isoformspecific ligand recognition and receptor recruitment
in the superfamily, J Biol. Chem. 285, 14806-
14814.
[7] Wrana, J. L., Attisano, L., Cárcamo, J., Zentella,
A., Doody, J., Laiho, M ,.Wang, X.-F., Massague,
J. (1992) TGFβ signals through a heteromeric
protein kinase receptor complex, Cell. 71, 1003-
1014.
[8] Groppe, J., Hinck, C. S., Samavarchi-Tehrani,
P., Zubieta, C., Schuermann, J. P., Taylor, A. B.,
Schwarz, P. M., Wrana, J. L., Hinck, A. P. (2008)
Cooperative assembly of TGF-β superfamily
signaling complexes is mediated by two disparate
mechanisms and distinct modes of receptor binding,
Mol. cell. 29, 157-168.
[9] Wrana, J. L., Attisano, L., Wieser, R., Ventura,
F., Massagué, J. (1994) Mechanism of activation of
the TGF-β receptor, Nature. 370, 341-347.
[10] Moustakas, A., Lin, H., Henis, Y. I.,
Plamondon, J. e., O'Connor-McCourt, M. &
Lodish, H. (1993) The transforming growth factor
beta receptors types I, II, and III form heterooligomeric complexes in the presence of ligand, J
Biol. Chem. 268, 22215-22218.
[11] Clark, D. A., Flanders, K. C., Hirte, H., Dasch,
J. R., Coker, R., McAnulty, R. J., Laurent, G. J.
(1995) Characterization of murine pregnancy
decidua transforming growth factor beta. I.
Transforming growth factor beta 2-like molecules
of unusual molecular size released in bioactive
form, Biol. Reprod. 52, 1380-1388.
[12] Schlunegger, M. P., Grütter, M. G. (1992) An
unusual feature revealed by the crystal structure at
2.2 Å resolution of human transforming growth fact
or-β2, Nature. 358, 430-434.
[13] Sun, P. D., Davies, D. R. (1995) The cystineknot growth-factor superfamily, Annu. Rev.
Biophys. Biomol. Struct. 24, 269-292.
[14] Mueller, T. D., Nickel, J. (2012) Promiscuity
and specificity in BMP receptor activation, FEBS
lett. 586, 1846-1859.