A heterologous immunoglobulin chain recombinant carries a distinct site for dinitrophenyl and obeys the common hapten binding mechanism
- PMID: 6794603
- DOI: 10.1021/bi00520a030
A heterologous immunoglobulin chain recombinant carries a distinct site for dinitrophenyl and obeys the common hapten binding mechanism
Abstract
A heterologous recombinant of the immunoglobulin alpha heavy chain derived from MOPC-460 and the lambda light chain from MOPC-315 was prepared. This H460L315 hybrid binds N epsilon-(2,4-dinitrophenyl)-L-lysine (DNPL) with an affinity of 1.6 X 10(4) M-1 (7 degrees C). This Ig-hapten complex exhibits an absorption spectrum which is different from those observed for each of its parent-DNPL complexes. Very small quenching is caused in the intrinsic fluorescence of the hybrid upon hapten binding, as contrasted by the large quenching of the parent molecules. Chemical relaxation kinetic measurements show that H460L315 exists in solution in two conformations which exchange with a relaxation time of 20 ms (7 degrees C). This transition is accompanied by a change in the fluorescence quantum yield of the protein. DNPL binds to both conformers at comparable fast, though not diffusion-controlled, rates. The equilibrium between the two conformers is shifted upon hapten binding, and the two complexes exchange at a faster rate than the free protein conformers. Thus H460L315 carries a new binding site for DNPL but follows the common mechanism of hapten binding as that observed for other immunoglobulins. These properties of the hybrid should be closely related to the interactions between its constituting domains.
Similar articles
-
A common mechanism of hapten binding to immunoglobulins and their heterologous chain recombinants.Biochemistry. 1980 Jun 10;19(12):2790-5. doi: 10.1021/bi00553a039. Biochemistry. 1980. PMID: 6772205
-
Kinetic evidence for hapten-induced conformational transition in immunoglobin MOPC 460.Proc Natl Acad Sci U S A. 1976 Oct;73(10):3549-53. doi: 10.1073/pnas.73.10.3549. Proc Natl Acad Sci U S A. 1976. PMID: 1068466 Free PMC article.
-
Dimerization kinetics of the IgE-class antibodies by divalent haptens. I. The Fab-hapten interactions.Biophys J. 1992 Aug;63(2):551-62. doi: 10.1016/S0006-3495(92)81609-0. Biophys J. 1992. PMID: 1420897 Free PMC article.
-
Binding of 2,4-dinitrophenyl derivatives by the light chain dimer obtained from immunoglobulin A produced by MOPC-315 mouse myeloma.Biochemistry. 1976 Jun 29;15(13):2785-90. doi: 10.1021/bi00658a013. Biochemistry. 1976. PMID: 820373
-
A functional examination of hapten-binding derivatives from a murine myeloma protein with immunoglobulin features.Proc Natl Acad Sci U S A. 1974 May;71(5):1940-4. doi: 10.1073/pnas.71.5.1940. Proc Natl Acad Sci U S A. 1974. PMID: 4525470 Free PMC article.
Cited by
-
The TCR binding site does move.Proc Natl Acad Sci U S A. 2007 Oct 16;104(42):16398-9. doi: 10.1073/pnas.0708462104. Epub 2007 Oct 9. Proc Natl Acad Sci U S A. 2007. PMID: 17925433 Free PMC article. No abstract available.
-
Structural invariants of antigen binding: comparison of immunoglobulin VL-VH and VL-VL domain dimers.Proc Natl Acad Sci U S A. 1985 Jul;82(14):4592-6. doi: 10.1073/pnas.82.14.4592. Proc Natl Acad Sci U S A. 1985. PMID: 3927286 Free PMC article.
-
What else can the immune system recognize?Proc Natl Acad Sci U S A. 1998 Sep 29;95(20):11509-10. doi: 10.1073/pnas.95.20.11509. Proc Natl Acad Sci U S A. 1998. PMID: 9751695 Free PMC article. Review. No abstract available.
-
Bound water molecules and conformational stabilization help mediate an antigen-antibody association.Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):1089-93. doi: 10.1073/pnas.91.3.1089. Proc Natl Acad Sci U S A. 1994. PMID: 8302837 Free PMC article.