Generation and Use of Recombinant Galectins.
galectin
galectin stability
galectin-1 alkylation using iodoacetamide
galectin-4
galectin-7
galectin-9 purification using Tris buffer
lactosyl-sepharose chromatography
stable and active galectin-9
Journal
Current protocols
ISSN: 2691-1299
Titre abrégé: Curr Protoc
Pays: United States
ID NLM: 101773894
Informations de publication
Date de publication:
Mar 2021
Mar 2021
Historique:
entrez:
3
3
2021
pubmed:
4
3
2021
medline:
22
6
2021
Statut:
ppublish
Résumé
Galectins are soluble carbohydrate binding proteins that can bind β-galactose-containing glycoconjugates by means of a conserved carbohydrate recognition domain (CRD). In mammalian systems, galectins have been shown to mediate very important roles in innate and adaptive immunity as well as facilitating host-pathogen relationships. Many of these studies have relied on purified recombinant galectins to uncover key features of galectin biology. A major limitation to this approach is that certain recombinant galectins purified using standard protocols are easily susceptible to loss of glycan-binding activity. As a result, biochemical studies that employ recombinant galectins can be misleading if the overall activity of a galectin remains unknown in a given assay condition. This article examines fundamental considerations when purifying galectins by lactosyl-sepharose and nickel-NTA affinity chromatography using human galectin-4N and -7 as examples, respectively. As other approaches are also commonly applied to galectin purification, we also discuss alternative strategies to galectin purification, using human galectin-1 and -9 as examples. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Purification of galectins using lactosyl-sepharose affinity chromatography Basic Protocol 2: Purification of human galectin-7 using a nickel-NTA affinity chromatography column Alternate Protocol 1: Iodoacetamide alkylation of free sulfhydryls on galectin-1 Alternate Protocol 2: Purification of human galectin-9 using lactosyl-sepharose column chromatography.
Substances chimiques
Carbohydrates
0
Galectins
0
Galactose
X2RN3Q8DNE
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e63Subventions
Organisme : NIH HHS
ID : DP5 OD019892
Pays : United States
Organisme : NHLBI NIH HHS
ID : T32 HL066987
Pays : United States
Organisme : NCI NIH HHS
ID : U01 CA242109
Pays : United States
Organisme : NIH HHS
ID : U01 CA242109
Pays : United States
Commentaires et corrections
Type : ErratumIn
Informations de copyright
© 2021 Wiley Periodicals LLC.
Références
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