Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 2000 Oct;79(4):2150–2154. doi: 10.1016/S0006-3495(00)76462-9

The unfolding/denaturation of immunogammaglobulin of isotype 2b and its F(ab) and F(c) fragments.

A W Vermeer 1, W Norde 1, A van Amerongen 1
PMCID: PMC1301104  PMID: 11023918

Abstract

The unfolding and further denaturation of IgG and its F(ab) and F(c) fragments were studied both on a macroscopic and molecular level, using differential scanning calorimetry and circular dichroism spectroscopy, respectively. It was shown that the structural integrity of the F(ab) and F(c) units was retained after fragmentation of the IgG. The F(ab) fragment denatured at approximately 61 degrees C and the F(c) fragment at 71 degrees C. The structural transitions observed in the whole IgG is the sum effect of those determined for the isolated F(ab) and F(c) fragments.

Full Text

The Full Text of this article is available as a PDF (83.4 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beaven M. A., Metzger H. Signal transduction by Fc receptors: the Fc epsilon RI case. Immunol Today. 1993 May;14(5):222–226. doi: 10.1016/0167-5699(93)90167-j. [DOI] [PubMed] [Google Scholar]
  2. Brandts J. F., Hu C. Q., Lin L. N., Mos M. T. A simple model for proteins with interacting domains. Applications to scanning calorimetry data. Biochemistry. 1989 Oct 17;28(21):8588–8596. doi: 10.1021/bi00447a048. [DOI] [PubMed] [Google Scholar]
  3. Brody T. Multistep denaturation and hierarchy of disulfide bond cleavage of a monoclonal antibody. Anal Biochem. 1997 May 1;247(2):247–256. doi: 10.1006/abio.1997.2062. [DOI] [PubMed] [Google Scholar]
  4. Burmeister W. P., Huber A. H., Bjorkman P. J. Crystal structure of the complex of rat neonatal Fc receptor with Fc. Nature. 1994 Nov 24;372(6504):379–383. doi: 10.1038/372379a0. [DOI] [PubMed] [Google Scholar]
  5. Chothia C., Novotný J., Bruccoleri R., Karplus M. Domain association in immunoglobulin molecules. The packing of variable domains. J Mol Biol. 1985 Dec 5;186(3):651–663. doi: 10.1016/0022-2836(85)90137-8. [DOI] [PubMed] [Google Scholar]
  6. Demignot S., Garnett M. C., Baldwin R. W. Mouse IgG2b monoclonal antibody fragmentation. Preparation and purification of Fab, Fc and Fab/c fragments. J Immunol Methods. 1989 Jul 26;121(2):209–217. doi: 10.1016/0022-1759(89)90162-2. [DOI] [PubMed] [Google Scholar]
  7. Goto Y., Ichimura N., Hamaguchi K. Effects of ammonium sulfate on the unfolding and refolding of the variable and constant fragments of an immunoglobulin light chain. Biochemistry. 1988 Mar 8;27(5):1670–1677. doi: 10.1021/bi00405a043. [DOI] [PubMed] [Google Scholar]
  8. Harrison P. T., Davis W., Norman J. C., Hockaday A. R., Allen J. M. Binding of monomeric immunoglobulin G triggers Fc gamma RI-mediated endocytosis. J Biol Chem. 1994 Sep 30;269(39):24396–24402. [PubMed] [Google Scholar]
  9. Lilie H., Buchner J. Domain interactions stabilize the alternatively folded state of an antibody Fab fragment. FEBS Lett. 1995 Mar 27;362(1):43–46. doi: 10.1016/0014-5793(95)00203-l. [DOI] [PubMed] [Google Scholar]
  10. Martsev S. P., Kravchuk Z. I., Vlasov A. P. Large increase in thermal stability of the CH2 domain of rabbit IgG after acid treatment as evidenced by differential scanning calorimetry. Immunol Lett. 1994 Dec;43(3):149–152. doi: 10.1016/0165-2478(94)90215-1. [DOI] [PubMed] [Google Scholar]
  11. Mian I. S., Bradwell A. R., Olson A. J. Structure, function and properties of antibody binding sites. J Mol Biol. 1991 Jan 5;217(1):133–151. doi: 10.1016/0022-2836(91)90617-f. [DOI] [PubMed] [Google Scholar]
  12. Miller S. Protein-protein recognition and the association of immunoglobulin constant domains. J Mol Biol. 1990 Dec 20;216(4):965–973. doi: 10.1016/S0022-2836(99)80014-X. [DOI] [PubMed] [Google Scholar]
  13. Oi V. T., Vuong T. M., Hardy R., Reidler J., Dangle J., Herzenberg L. A., Stryer L. Correlation between segmental flexibility and effector function of antibodies. Nature. 1984 Jan 12;307(5947):136–140. doi: 10.1038/307136a0. [DOI] [PubMed] [Google Scholar]
  14. Padlan E. A. Anatomy of the antibody molecule. Mol Immunol. 1994 Feb;31(3):169–217. doi: 10.1016/0161-5890(94)90001-9. [DOI] [PubMed] [Google Scholar]
  15. Raghavan M., Bjorkman P. J. Fc receptors and their interactions with immunoglobulins. Annu Rev Cell Dev Biol. 1996;12:181–220. doi: 10.1146/annurev.cellbio.12.1.181. [DOI] [PubMed] [Google Scholar]
  16. Ravetch J. V., Kinet J. P. Fc receptors. Annu Rev Immunol. 1991;9:457–492. doi: 10.1146/annurev.iy.09.040191.002325. [DOI] [PubMed] [Google Scholar]
  17. Tischenko V. M., Zav'yalov V. P., Medgyesi G. A., Potekhin S. A., Privalov P. L. A thermodynamic study of cooperative structures in rabbit immunoglobulin G. Eur J Biochem. 1982 Sep 1;126(3):517–521. doi: 10.1111/j.1432-1033.1982.tb06811.x. [DOI] [PubMed] [Google Scholar]
  18. Underdown B. J., Schiff J. M. Immunoglobulin A: strategic defense initiative at the mucosal surface. Annu Rev Immunol. 1986;4:389–417. doi: 10.1146/annurev.iy.04.040186.002133. [DOI] [PubMed] [Google Scholar]
  19. Vermeer A. W., Bremer M. G., Norde W. Structural changes of IgG induced by heat treatment and by adsorption onto a hydrophobic Teflon surface studied by circular dichroism spectroscopy. Biochim Biophys Acta. 1998 Sep 16;1425(1):1–12. doi: 10.1016/s0304-4165(98)00048-8. [DOI] [PubMed] [Google Scholar]
  20. Vermeer A. W., Norde W. The thermal stability of immunoglobulin: unfolding and aggregation of a multi-domain protein. Biophys J. 2000 Jan;78(1):394–404. doi: 10.1016/S0006-3495(00)76602-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Vermeer Arnoldus W. P., Norde Willem. CD Spectroscopy of Proteins Adsorbed at Flat Hydrophilic Quartz and Hydrophobic Teflon Surfaces. J Colloid Interface Sci. 2000 May 15;225(2):394–397. doi: 10.1006/jcis.2000.6769. [DOI] [PubMed] [Google Scholar]
  22. van de Winkel J. G., Capel P. J. Human IgG Fc receptor heterogeneity: molecular aspects and clinical implications. Immunol Today. 1993 May;14(5):215–221. doi: 10.1016/0167-5699(93)90166-I. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES