Imparting Exquisite Specificity to Peanut Agglutinin for the Tumor-associated Thomsen-Friedenreich Antigen by Redesign of Its Combining Site

Sharma, Vivek and Vijayan, M and Surolia, Avadhesha (1996) Imparting Exquisite Specificity to Peanut Agglutinin for the Tumor-associated Thomsen-Friedenreich Antigen by Redesign of Its Combining Site. In: The Journal of Biological Chemistry, 271 (35). pp. 21209-21213.

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Abstract

Lectins from legumes constitute one of the most thoroughly studied families of proteins, yet the absence of a rigorous framework to explain their carbohydrate binding specificities appears to have prevented a rational approach to alter their ligand binding activity. Studies reported here deal with the redesign of the recognition propensity of peanut agglutinin (PNA), an important member of the family. PNA is extensively used as a tool for recognition of the tumor-associated Thomsen-Friedenrich antigen (T-antigen; Galb1–3GalNAc) on the surfaces of malignant cells and immature thymocytes. PNA also recognizes N-acetyllactosamine (LacNAc;Galb1–4GlcNAc), which is present at the termini of several cell-surface glycoproteins. The crystal structure of the PNA-lactose complex revealed, in addition to the expected interactions with the residues constituting the binding site, the presence of leucine 212 at a position close enough to be in steric contact with the acetamido group on LacNAc. We report here two leucine mutants, one to asparagine (L212N) and the other to alanine(L212A),that exhibit distinct preference for T-antigen and N-acetyllactosamine, respectively. Carbohydrate binding studies reveal that mutant L212N does not recognize LacNAc at high concentrations, thus making it an exquisitely specific cell-surface marker compared with its wild-type counterpart.

Item Type:	Journal Article
Publication:	The Journal of Biological Chemistry
Publisher:	The American Society for Biochemistry and Molecular Biology, Inc.
Additional Information:	The copyright belongs to The American Society for Biochemistry and Molecular Biology, Inc.
Keywords:	agglutinin;n acetyllactosamine;lectins;carbohydrate binding
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited:	10 Aug 2004
Last Modified:	19 Sep 2010 04:14
URI:	http://eprints.iisc.ac.in/id/eprint/1245

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