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Biomol NMR Assign. 2016 Apr;10(1):25-9. doi: 10.1007/s12104-015-9630-2. Epub 2015 Aug 15.

Near-complete 1H, 13C, 15N resonance assignments of dimethylsulfoxide-denatured TGFBIp FAS1-4 A546T.

Author information

1
Department of Chemistry, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark.
2
Max-Plank Institute, Am Fassberg 11, 37077, Göttingen, Germany.
3
Department of Molecular Biology and Genetics, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark.
4
Department of Chemistry, Cambridge University, Lensfield Road, Cambridge, CB2 1EW, UK.
5
Department of Chemistry, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark. fmulder@chem.au.dk.

Abstract

The transforming growth factor beta induced protein (TGFBIp) is a major protein component of the human cornea. Mutations occurring in TGFBIp may cause corneal dystrophies, which ultimately lead to loss of vision. The majority of the disease-causing mutations are located in the C-terminal domain of TGFBIp, referred as the fourth fascilin-1 (FAS1-4) domain. In the present study the FAS1-4 Ala546Thr, a mutation that causes lattice corneal dystrophy, was investigated in dimethylsulfoxide using liquid-state NMR spectroscopy, to enable H/D exchange strategies for identification of the core formed in mature fibrils. Isotope-labeled fibrillated FAS1-4 A546T was dissolved in a ternary mixture 95/4/1 v/v/v% dimethylsulfoxide/water/trifluoroacetic acid, to obtain and assign a reference 2D (1)H-(15)N HSQC spectrum for the H/D exchange analysis. Here, we report the near-complete assignments of backbone and aliphatic side chain (1)H, (13)C and (15)N resonances for unfolded FAS1-4 A546T at 25 °C.

KEYWORDS:

Corneal dystrophy; DMSO; FAS1-4 domain; H/D-exchange NMR spectroscopy; TGFBIp

PMID:
26275916
DOI:
10.1007/s12104-015-9630-2
[Indexed for MEDLINE]

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