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Backbone and side-chain 1H, 13C and 15N resonance assignments of LEN, a human immunoglobulin κIV light-chain variable domain

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Abstract

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.

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References

  • Baden EM, Owen BA, Peterson FC, Volkman BF, Ramirez-Alvarado M, Thompson JR (2008) Altered dimer interface decreases stability in an amyloidogenic protein. J Biol Chem 283:15853–15860

    Article  Google Scholar 

  • Bourne PC, Ramsland PA, Shan L, Fan ZC, DeWitt CR, Shultz BB, Terzyan SS, Moomaw CR, Slaughter CA, Guddat LW, Edmundson AB (2002) Three-dimensional structure of an immunoglobulin light-chain dimer with amyloidogenic properties. Acta Crystallogr D 58:815–823

    Article  Google Scholar 

  • Cai ML, Huang Y, Sakaguchi K, Clore GM, Gronenborn AM, Craigie R (1998) An efficient and cost-effective isotope labeling protocol for proteins expressed in Escherichia coli. J Biomol NMR 11:97–102

    Article  Google Scholar 

  • Cavanagh J, Fairbrother WJ, Palmer AG, Skelton NJ (1996) Protein NMR Spectroscopy: principles and practice. Academic Press, San Diego

    Google Scholar 

  • Chiti F, Dobson CM (2006) Protein misfolding, functional amyloid, and human disease. Annu Rev Biochem 75:333–366

    Article  Google Scholar 

  • Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6:277–293

    Article  Google 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):3–4952

    Google Scholar 

  • Falk RH, Comenzo RL, Skinner M (1997) The systemic amyloidoses. N Engl J Med 337:898–909

    Article  Google Scholar 

  • Farrow NA, Muhandiram R, Singer AU, Pascal SM, Kay CM, Gish G, Shoelson SE, Pawson T, Forman-Kay JD, Kay LE (1994) Backbone dynamics of a free and a phosphopeptide-complexed Src homology 2 domain studied by 15N NMR relaxation. Biochemistry 33:5984–6003

    Article  Google Scholar 

  • Goddard TD, Kneller DG (1993) SPARKY 3, University of California, San Francisco

  • Gutierrez-Gonzalez LH, Muresanu L, del Pozo-Yauner L, Sanchez R, Guereca L, Becerril B, Lucke C (2007) 1H, 13C and 15N resonance assignment of 6aJL2(R25G), a highly fibrillogenic λVI light chain variable domain. Biomol NMR Assign 1:159–161

    Article  Google Scholar 

  • Huang DB, Chang CH, Ainsworth C, Brunger AT, Eulitz M, Solomon A, Stevens FJ, Schiffer M (1994) Comparison of two homologous proteins: structural origin of altered domain interactions in immunoglobulin light-chain dimers. Biochemistry 33:14848–14857

    Article  Google Scholar 

  • Huang DB, Chang CH, Ainsworth C, Johnson G, Solomon A, Stevens FJ, Schiffer M (1997) Variable domain structure of κIV human light chain Len: high homology to the murine light chain McPC603. Mol Immunol 34:1291–1301

    Article  Google Scholar 

  • Hurle MR, Helms LR, Li L, Chan W, Wetzel R (1994) A role for destabilizing amino acid replacements in light-chain amyloidosis. Proc Natl Acad Sci USA 91:5446–5450

    Article  ADS  Google 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, DC

    Google Scholar 

  • Khurana R, Gillespie JR, Talapatra A, Minert LJ, Ionescu-Zanetti C, Millett I, Fink AL (2001) Partially folded intermediates as critical precursors of light chain amyloid fibrils and amorphous aggregates. Biochemistry 40:3525–3535

    Article  Google Scholar 

  • Kim YS, Wall JS, Meyer J, Murphy C, Randolph TW, Manning MC, Solomon A, Carpenter JF (2000) Thermodynamic modulation of light chain amyloid fibril formation. J Biol Chem 275:1570–1574

    Article  Google Scholar 

  • Kyle RA, Gertz MA (1995) Primary systemic amyloidosis: clinical and laboratory features in 474 cases. Semin Hematol 32:45–59

    Google Scholar 

  • Palmer AG, Rance M, Wright PE (1991) Intramolecular motions of a zinc finger DNA-binding domain from Xfin characterized by proton-detected natural abundance 13C heteronuclear NMR spectroscopy. J Am Chem Soc 113:4371–4380

    Article  Google 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 8:509–517

    Google Scholar 

  • Schormann N, Murrell JR, Liepnieks JJ, Benson MD (1995) Tertiary structure of an amyloid immunoglobulin light chain protein: a proposed model for amyloid fibril formation. Proc Natl Acad Sci USA 92:9490–9494

    Article  ADS  Google Scholar 

  • Solomon A, Weiss DT (1995) Protein and host factors implicated in the pathogenesis of light chain amyloidosis (AL amyloidosis). Amyloid 2:269–279

    Article  Google Scholar 

  • Souillac PO, Uversky VN, Millett IS, Khurana R, Doniach S, Fink AL (2002) Effect of association state and conformational stability on the kinetics of immunoglobulin light chain amyloid fibril formation at physiological pH. J Biol Chem 277:12657–12665

    Article  Google Scholar 

  • Souillac PO, Uversky VN, Fink AL (2003) Structural transformations of oligomeric intermediates in the fibrillation of the immunoglobulin light chain LEN. Biochemistry 42:8094–8104

    Article  Google 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 17:323–331

    Article  Google 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 4:421–432

    Article  Google Scholar 

  • Yamazaki T, Lee W, Arrowsmith CH, Muhandiram DR, Kay LE (1994) A suite of triple resonance NMR experiments for the backbone assignment of 15N, 13C, 2H labeled proteins with high sensitivity. J Am Chem Soc 116:11655–11666

    Article  Google Scholar 

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Acknowledgments

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.

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Authors and Affiliations

  1. Department of Chemistry, The Ohio State University, 100 West 18th Avenue, Columbus, OH, 43210, USA

    Sujoy Mukherjee, Simon P. Pondaven, Nicole Höfer & Christopher P. Jaroniec

  2. Department of Chemical and Environmental Sciences, University of Limerick, Limerick, Ireland

    Nicole Höfer

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  1. Sujoy Mukherjee
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  2. Simon P. Pondaven
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  3. Nicole Höfer
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  4. Christopher P. Jaroniec
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Correspondence to Christopher P. Jaroniec.

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Mukherjee, S., Pondaven, S.P., Höfer, N. et al. Backbone and side-chain 1H, 13C and 15N resonance assignments of LEN, a human immunoglobulin κIV light-chain variable domain. Biomol NMR Assign 3, 255–259 (2009). https://doi.org/10.1007/s12104-009-9188-y

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