The use of the Congo red-related dye DBACR to recognize the heavy chain-derived abnormality of myeloma immunoglobulins

Abstract.IntroductionThe aim of this study was to differentiate heavy and light chain-derived instability of monoclonal myeloma immunoglobulins by complexation of matched supramolecular dyes. These are composed of several micellar pieces of self-assembled dye molecules which may penetrate the protein interior of the binding locus with polypeptide chains. These dyes were used to elicit, by precipitation, the postulated higher aggregation tendency of the heavy chain derived from its higher hydrophobicity.Materials and MethodsAgarose gel electrophoresis was used to create conditions for dye complexation and to reveal the precipitation.ResultsCongo red derivatives with aromatic ring substitutes, BACR and DBACR, of increased penetrating capability were chosen to provoke the precipitation of abnormal immunoglobulins by displacing association-prone polypeptide chains from the protein interior.ConclusionsThe results of this study confirm the heavy chain-related propensity of some monoclonal immunoglobulins to aggregate and precipitate. The simplicity of the technique may improve clinical diagnosis and facilitate predictions of disease complications.

[1]  I. Roterman,et al.  The structural abnormality of myeloma immunoglobulins tested by Congo red binding. , 2003, Medical science monitor : international medical journal of experimental and clinical research.

[2]  J. Stachura,et al.  Generalized crystal-storing histiocytosis as a presentation of multiple myeloma: a case with a possible pro-aggregation defect in the immunoglobulin heavy chain , 2003, Virchows Archiv.

[3]  C Chothia,et al.  Structural determinants in the sequences of immunoglobulin variable domain. , 1998, Journal of molecular biology.

[4]  Leszek Konieczny,et al.  The structure and protein binding of amyloid-specific dye reagents. , 2003, Acta biochimica Polonica.

[5]  D. Pressman,et al.  SPECIFIC COMBINATION OF H AND L CHAINS OF RABBIT γ-GLOBULINS , 1964 .

[6]  I. Roterman,et al.  Supramolecular ligands: monomer structure and protein ligation capability. , 1998, Biochimie.

[7]  D. Pressman,et al.  SPECIFIC COMBINATION OF H AND L CHAINS OF RABBIT GAMMA-GLOBULINS. , 1964, Proceedings of the National Academy of Sciences of the United States of America.

[8]  P. Adams,et al.  Structural basis of light chain amyloidogenicity: comparison of the thermodynamic properties, fibrillogenic potential and tertiary structural features of four Vλ6 proteins , 2004, Journal of molecular recognition : JMR.

[9]  Y. Argon,et al.  Inhibition of amyloid fiber assembly by both BiP and its target peptide. , 2000, Immunity.

[10]  I Roterman,et al.  Congo red-stabilized intermediates in the lambda light chain transition from native to molten state. , 1996, Biochimie.

[11]  Andreas Plückthun,et al.  Biophysical properties of human antibody variable domains. , 2003, Journal of molecular biology.

[12]  I. Roterman,et al.  The melting of native domain structure in effector activation of IgG studied by using congo red as a specific probe. , 1994, Journal of physiology and pharmacology : an official journal of the Polish Physiological Society.

[13]  M. Karplus,et al.  CHARMM: A program for macromolecular energy, minimization, and dynamics calculations , 1983 .

[14]  I. Roterman,et al.  Analysis of correlated domain motions in IgG light chain reveals possible mechanisms of immunological signal transduction , 2005, Proteins.

[15]  S. Muyldermans,et al.  Single domain antibodies: comparison of camel VH and camelised human VH domains. , 1999, Journal of immunological methods.

[16]  Leszek Konieczny,et al.  Instability of monoclonal myeloma protein may be identified as susceptibility to penetration and binding by newly synthesized Congo red derivatives. , 2004, Biochimie.

[17]  G. Merlini,et al.  Protein Aggregation , 2001, Clinical chemistry and laboratory medicine.

[18]  S. Cohen Properties of the Separated Chains of Human γ-Globulin , 1963, Nature.