Purification and Biochemical Characterization of Selected F-Type Lectins

Methods Mol Biol. 2020:2132:241-255. doi: 10.1007/978-1-0716-0430-4_24.

Abstract

The purification of fucose-binding lectins from the liver of striped bass (Morone saxatilis), a teleost fish, and the identification of a novel lectin sequence motif led to the identification of a new family of lectins, the F-type lectins (FTLs) (see overview of the FTL family in Chapter 23 ). Isolation and purification of these proteins from liver extracts of striped bass was accomplished by affinity chromatography and size exclusion, and their identification as FTLs, by direct Edman sequencing, and protein, transcript, and gene sequence analysis. The development of specific antibodies against the M. saxatilis FTL provided an additional tool for the identification of FTLs. These methods have been successfully used for the purification of the FTL family members from tissues and body fluids of various animal species. Production and characterization of FTLs has been facilitated by the expression of the recombinant proteins. In this chapter, the biochemical characterization of FTLs is focused on the analysis of their carbohydrate specificity.

Keywords: Affinity and size-exclusion chromatography; Carbohydrate specificity; F-type lectin sequence motif; Recombinant F-type lectin; Sequence analysis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Bass / genetics
  • Bass / metabolism*
  • Chromatography, Affinity
  • Chromatography, Gel
  • Fish Proteins / metabolism
  • Gene Expression Profiling
  • Gene Expression Regulation
  • Lectins / genetics*
  • Lectins / isolation & purification*
  • Lectins / metabolism
  • Liver / metabolism*
  • Multigene Family
  • Recombinant Proteins / metabolism

Substances

  • Fish Proteins
  • Lectins
  • Recombinant Proteins
  • fucose-binding lectin