Near-complete 1H, 13C, 15N resonance assignments of dimethylsulfoxide-denatured TGFBIp FAS1-4 A546T.
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.
Department of Chemistry, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark.|Max-Plank Institute, Am Fassberg 11, 37077, Göttingen, Germany.
Natalia V
NV
Kulminskaya
Department of Chemistry, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark.
Yuichi
Y
Yoshimura
Department of Molecular Biology and Genetics, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark.
Kasper
K
Runager
Department of Molecular Biology and Genetics, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark.
Charlotte S
CS
Sørensen
Department of Chemistry, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark.
Morten
M
Bjerring
Department of Molecular Biology and Genetics, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark.|Department of Chemistry, Cambridge University, Lensfield Road, Cambridge, CB2 1EW, UK.
Maria
M
Andreasen
Department of Molecular Biology and Genetics, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark.
Daniel E
DE
Otzen
Department of Molecular Biology and Genetics, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark.
Jan J
JJ
Enghild
Department of Chemistry, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark.
Niels Chr
NC
Nielsen
Department of Chemistry, Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, 8000, Aarhus C, Denmark. fmulder@chem.au.dk.
Frans A A
FA
Mulder
D000595:Amino Acid Sequence; D002247:Carbon Isotopes; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D016326:Extracellular Matrix Proteins / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D050505:Mutant Proteins / Q000737:chemistry* / Q000378:metabolism*; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D011489:Protein Denaturation / Q000187:drug effects*; D000072417:Protein Domains; D016212:Transforming Growth Factor beta / Q000737:chemistry* / Q000378:metabolism*; D014316:Tritium; D000099222:betaIG-H3 Protein
D002247:Carbon Isotopes; D004121:Dimethyl Sulfoxide; D016326:Extracellular Matrix Proteins; D050505:Mutant Proteins; D009587:Nitrogen Isotopes; D016212:Transforming Growth Factor beta; D014316:Tritium; D000099222:betaIG-H3 Protein
Corneal dystrophy; DMSO; FAS1-4 domain; H/D-exchange NMR spectroscopy; TGFBIp