Multidimensional protein NMR: studies of lectin-carbohydrate interactions and of the hapten binding in one domain antibodies.

Project Details

Description

Project aiming at addressing structure and functions of proteins in two important areas of protein chemistry: sugar binding lectins and one-domain antibodies. The structures are addressed in aqueous solutions by multidimensional gradient pulsed NMR assisted by molecular modelling. Lectins are important in the antifungal immune response of plants, while one-domain antibodies have recently been discovered for the first time in mammalians (camelidae) and have important potential s in biotechnological applications. The mechanisms of the functions of these proteins remain largely to be unravelled. The project is presently being extended, in the frame of a concerted action and a interuniversity attraction pole to other systems, among which an arsenate binding protein.This project, to be conducted in close collaboration with the units ULTR and ALBI of the VUB, aims at investigating structural features in solution of some practically important proteins, while applying and setting up multidimensional, homo- and heteronuclear, gradient assisted NMR techniques. Two types of questions will be addressed. First, the structure of a number of lectins, small proteins without enzymatic activity binding sugars, will be studied. Lectins play a role in the antifungal response of plants. The exact nature of their conformations, their interactions with different types of solvation water molecules and their binding with carbohydrates remain to be elucidated. Understanding such host-guest interactions is of major importance in pharmacodynamics. The NMR studies planned will allow to precize the dynamic behaviour of lectins and their substrates in conditions mimicking as well as possible the intracellular medium. The lectins to be examined originate from Urtica Dioica and Amaranthus Caudatus. Second, the structural features of one-domain antibodies will be investigated. These are of importance in biotechnological applications, especially in strategies aiming at simplifying their genetic expression in bacteria. The NMR-studies will be conducted with 13C and lSN enriched one-domain antibodies of animal origin. The concrete quesdons to be addressed are: 1. the 3D structure of their VH domains so as to characterize their water structure and the dynamics of their CDR loops; 2. a ligand binding involving some haptens or peptide epitopes from the Iysozyme model system. 3. the hinge peptides, either as isolated peptide or as part of a larger Vh-Ch2 construction.
Acronym1940440030
StatusFinished
Effective start/end date1/01/9431/12/97

Keywords

  • HNMR
  • biology

Flemish discipline codes

  • Biological sciences