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Purification and Biochemical Characterization of Selected F-Type Lectins

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Lectin Purification and Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2132))

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.

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Abbreviations

BSA:

Bovine serum albumin

BSM:

Bovine submaxillary mucin

CBB:

Coomassie Brilliant Blue

ELISA:

Enzyme-linked immunosorbent assay

FBD:

Fucose-binding domains

FTL:

F-type lectin

Igs:

Immunoglobulins

IPTG:

Isopropyl β-D-thiogalactoside

MsaFBP32:

M. saxatilis fucose-binding protein 32 kDa

NCFG:

National Center for Functional Glycomics

OD:

Optical density

OSM:

Ovine submaxillary mucin

PBS:

Phosphate-buffered saline

PMSF:

Phenylmethylsulfonyl fluoride

PSM:

Porcine stomach mucin

RBC:

Red blood cells

RP-HPLC:

Reverse phase-HPLC

SEC:

Size exclusion chromatography

TFA:

Trifluoroacetic acid

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Acknowledgments

The author’s research reviewed herein was supported by Grants IOS 1050518, IOB-0618409, MCB 0077928, and IOS-0822257 from the National Science Foundation, and Grant R01GM070589 from the National Institutes of Health to G.R.V.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, University of Maryland School of Medicine, UMB, and Institute of Marine and Environmental Technology, Columbus Center, Baltimore, MD, USA

    Chiguang Feng & Gerardo R. Vasta

Authors
  1. Chiguang Feng
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  2. Gerardo R. Vasta
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Correspondence to Gerardo R. Vasta .

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Editors and Affiliations

  1. Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan

    Jun Hirabayashi

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Feng, C., Vasta, G.R. (2020). Purification and Biochemical Characterization of Selected F-Type Lectins. In: Hirabayashi, J. (eds) Lectin Purification and Analysis. Methods in Molecular Biology, vol 2132. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0430-4_24

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  • DOI: https://doi.org/10.1007/978-1-0716-0430-4_24

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0429-8

  • Online ISBN: 978-1-0716-0430-4

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