Cholera toxin binding affinity and specificity for gangliosides determined by surface plasmon resonance
- Univ. of California, Berkeley, CA (United States)
The present study determines the affinity of cholera toxin for the ganglioside series GM1, GM2, GM3, GD1A, GD1B, GT1B, asialo GM1, globotriosyl ceramide, and lactosyl ceramide using real time biospecific interaction analysis (surface plasmon resonance, SPR). SPR shows that cholera toxin preferably binds to gangliosides in the following sequence: GM1 > GM2 > GD1A > GM3 > GT1B > GD1B > asialo-GM1. The measured binding affinity of cholera toxin for the ganglioside sequence ranges from 4.61 {times} 10{sup {minus}12} M for GM1 to 1.88 {times} 10{sup {minus}10} M for asialo GM1. The picomolar values obtained by surface plasmon resonance are similar to K{sub d} values determined with whole-cell binding assays. Both whole-cell assays ans SPR measurements on synthetic membranes are higher than free solution measurements by several orders of magnitude. This difference may be caused by the effects of avidity and charged lipid head-groups, which may play a major role in the binding between cholera toxin, the receptor, and the membrane surface. The primary difference between free solution binding studies and surface plasmon resonance studies is that the latter technique is performed on surfaces resembling the cell membrane. Surface plasmon resonance has the further advantage of measuring apparent kinetic association and dissociation rates in real time, providing direct information about binding events at the membrane surface. 34 refs., 8 figs., 2 tabs.
- DOE Contract Number:
- AC03-76SF00098
- OSTI ID:
- 518242
- Journal Information:
- Biochemistry (Eaton), Journal Name: Biochemistry (Eaton) Journal Issue: 20 Vol. 35; ISSN 0006-2960; ISSN BICHAW
- Country of Publication:
- United States
- Language:
- English
Similar Records
Approaches in the study of ganglioside metabolism
Alteration of glycolipids in ras-transfected NIH 3T3 cells
A photoreactive derivative of radiolabeled GM1 ganglioside: Preparation and use to establish the involvement of specific proteins in GM1 uptake by human fibroblasts in culture
Journal Article
·
Sat Dec 31 23:00:00 EST 1983
· Adv. Exp. Med. Biol.; (United States)
·
OSTI ID:6349876
Alteration of glycolipids in ras-transfected NIH 3T3 cells
Journal Article
·
Tue Sep 01 00:00:00 EDT 1987
· Proc. Natl. Acad. Sci. U.S.A.; (United States)
·
OSTI ID:5294148
A photoreactive derivative of radiolabeled GM1 ganglioside: Preparation and use to establish the involvement of specific proteins in GM1 uptake by human fibroblasts in culture
Journal Article
·
Mon Jan 09 23:00:00 EST 1989
· Biochemistry; (USA)
·
OSTI ID:5482341