Backbone and side-chain 1H, 13C and 15N resonance assignments of LEN, a human immunoglobulin κIV light-chain variable domain
1H, 13C and 15N resonance assignments are presented for a recombinant 114 amino acid human immunoglobulin (Ig) κIV light-chain variable domain (VL) LEN, which displays a high degree of sequence identity with another human Ig κIV VL, SMA. While SMA is highly amyloidogenic in vivo and in vitro and has been linked to the pathogenesis of light-chain amyloidosis, LEN is non-amyloidogenic in vivo and can be converted to the amyloid state only in vitro under destabilizing conditions. Measurements of longitudinal and transverse amide 15N relaxation rates confirm that, as expected, LEN is a dimer at physiological pH and typical concentrations used for NMR studies, and the analysis of secondary chemical shifts indicates that the protein has a high β-sheet content. These findings are consistent with previously published biophysical data and the high-resolution X-ray structure of LEN.
KeywordsImmunoglobulin light-chain Bence Jones protein Light-chain amyloidosis Protein misfolding NMR spectroscopy
This research was supported by a grant from the American Heart Association (0865410D) and a Young Investigator Award from Eli Lilly and Company to C.P.J. S.M. thanks the American Heart Association for a Postdoctoral Fellowship (09POST2220178). N.H. acknowledges support from grants provided by the Science Foundation Ireland (07/IN.1/B1836) and the National Institutes of Health (GM75915) to Dr. Martin Caffrey (University of Limerick). We thank Dr. Fred J. Stevens for the gift of the LEN plasmid.
- Cavanagh J, Fairbrother WJ, Palmer AG, Skelton NJ (1996) Protein NMR Spectroscopy: principles and practice. Academic Press, San DiegoGoogle Scholar
- Epp O, Lattman EE, Schiffer M, Huber R, Palm R (1975) The molecular structure of a dimer composed of the variable portions of the Bence-Jones protein REI refined at 2.0 Å resolution. Biochemistry 14(494):3–4952Google Scholar
- Goddard TD, Kneller DG (1993) SPARKY 3, University of California, San FranciscoGoogle Scholar
- Kabat EA, Wu TT, Perry HM, Gottesman KS, Foeller C (1991) Sequences of proteins of immunological interest, 5th edn. NIH publication No. 91-3242, U.S., Department of Health and Human Services, Washington, DCGoogle Scholar
- Kyle RA, Gertz MA (1995) Primary systemic amyloidosis: clinical and laboratory features in 474 cases. Semin Hematol 32:45–59Google Scholar
- Raffen R, Dieckman LJ, Szpunar M, Wunschl C, Pokkuluri PR, Dave P, Wilkins Stevens P, Cai X, Schiffer M, Stevens FJ (1999) Physicochemical consequences of amino acid variations that contribute to fibril formation by immunoglobulin light chains. Protein Sci 8:509–517Google Scholar
- Wall JS, Gupta V, Wilkerson M, Schell M, Loris R, Adams P, Solomon A, Stevens F, Dealwis C (2004) Structural basis of light chain amyloidogenicity: comparison of the thermodynamic properties, fibrillogenic potential and tertiary structural features of four Vλ6 proteins. J Mol Recognit 17:323–331CrossRefGoogle Scholar
- Wilkins Stevens P, Raffen R, Hanson DK, Deng YL, Berrios-Hammond M, Westholm FA, Murphy C, Eulitz M, Wetzel R, Solomon A, Schiffer M, Stevens FJ (1995) Recombinant immunoglobulin variable domains generated from synthetic genes provide a system for in vitro characterization of light-chain amyloid proteins. Protein Sci 4:421–432CrossRefGoogle Scholar