Vertebrate lectins, Comparison of properties of beta-galactoside-binding lectins from tissues of calf and chicken.

Briles, Eve Barak; Gregory, William F.; Fletcher, Paul L.; Kornfeld, Stuart

doi:10.1083/jcb.81.3.528

Cited by 131 publications

(31 citation statements)

References 9 publications

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“…It should be noted that previous studies had shown that Gal1 forms non-covalently associated dimers that exist in equilibrium with monomeric Gal1 [33–35]. Therefore, it was possible that the observed pull-down of TFII-I by GST-Gal1(N46D) can be accounted for by hybrid dimeric molecules (consisting of one GST-Gal1(N46D) and one wild-type Gal1 derived from NE) tethered to the glutathione beads via GST.…”

Section: Discussionmentioning

confidence: 99%

Dissociation of the carbohydrate-binding and splicing activities of galectin-1

Voss

Gray

Dickey

et al. 2008

Archives of Biochemistry and Biophysics

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Galectin-1 (Gal1) and galectin-3 (Gal3) are two members of a family of carbohydrate-binding proteins that are found in the nucleus and that participate in pre-mRNA splicing assayed in a cellfree system. When nuclear extracts (NE) of HeLa cells were subjected to adsorption on a fusion protein containing glutathione S-transferase (GST) and Gal3, the general transcription factor II-I (TFII-I) was identified by mass spectrometry as one of the polypeptides specifically bound. Lactose and other saccharide ligands of the galectins inhibited GST-Gal3 pull-down of TFII-I while nonbinding carbohydrates failed to yield the same effect. Similar results were also obtained using GSTGal1. Site-directed mutants of Gal1, expressed and purified as GST fusion proteins, were compared with the wild-type (WT) in three assays: (a) binding to asialofetuin-Sepharose as a measure of the carbohydrate-binding activity; (b) pull-down of TFII-I from NE; and (c) reconstitution of splicing in NE depleted of galectins as a test of the in vitro splicing activity. The binding of GST-Gal1(N46D) to asialofetuin-Sepharose was less than 10% of that observed for GST-Gal1(WT), indicating that the mutant was deficient in carbohydrate-binding activity. In contrast, both GST-Gal1(WT) and GSTGal1(N46D) were equally efficient in pull-down of TFII-I and in reconstitution of splicing activity in the galectin-depleted NE. Moreover, while the splicing activity of the wild-type protein can be inhibited by saccharide ligands, the carbohydrate-binding deficient mutant was insensitive to such inhibition. Together, all of the results suggest that the carbohydrate-binding and the splicing activities of Gal1 can be dissociated and therefore, saccharide-binding, per se, is not required for the splicing activity.

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Section: Discussionmentioning

confidence: 99%

Dissociation of the carbohydrate-binding and splicing activities of galectin-1

Voss

Gray

Dickey

et al. 2008

Archives of Biochemistry and Biophysics

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“…A vertebrate 0-galactoside lectin with Mr 29,000 under dissociating and reducing conditions has not been observed in rat (15), bovine (8,16), or human (15, 16) tissues prepared by methods like those used here, although it is detectable in the original preparation of bovine lectin (7) . CLL-1 (14-16) and electrolectin (34), related carbohydrate-binding proteins, also have subunit molecular weights in the range of 15,000 but no larger subunits were detected by gel electrophoresis under dissociating conditions.…”

Section: Endogenous Lectin In Elastic Fibersmentioning

confidence: 99%

“…A 0-galactoside-binding lectin has been identified in several mammalian tissues including bovine (7,8) and rat (9,15) lung. Like CLL-I, it is a dimer under nondissociating conditions.…”

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confidence: 99%

Endogenous mammalian lectin localized extracellularly in lung elastic fibers

Cerra¹

1984

The Journal of Cell Biology

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An affinity-purified antibody preparation raised against a beta- galactoside-binding lectin from bovine lung was used to localize a similar lectin in rat lung by immunofluorescence and by electron microscopy after on-grid staining visualized with colloidal gold conjugated second antibody. The endogenous mammalian lectin was found in smooth muscle cells and squamous alveolar epithelial (type I) cells and was concentrated extracellularly in elastic fibers of pulmonary parenchyma and blood vessels. The extracellular localization of this lectin suggests that it, like others, functions by interaction with extracellular glycoconjugates.

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“…Purification and Characterization of Bovine Heart dGal-1-Bovine heart dGal-1 was purified essentially as described (53,54) with some modifications. Briefly, frozen bovine heart was trimmed of fat, cut to small pieces, and minced for 15 s in an Osterizer blender using an S blade (low speed, shred; Sunbeam Products, Inc., Boca Raton, FL).…”

Section: Methodsmentioning

confidence: 99%

Dimeric Galectin-1 Binds with High Affinity to α2,3-Sialylated and Non-sialylated Terminal N-Acetyllactosamine Units on Surface-bound Extended Glycans

Leppänen

Stowell

Blixt

et al. 2005

Journal of Biological Chemistry

152

141

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Galectin-1 is a member of the galectin family of glycan-binding proteins and occurs as an ϳ29.5-kDa noncovalent homodimer (dGal-1) that is widely expressed in many tissues. Here, we report that human recombinant dGal-1 bound preferentially and with high affinity (apparent K d ϳ 2-4 M) to immobilized extended glycans containing terminal N-acetyllactosamine (LN; Gal␤1-4GlcNAc) sequences on poly-N-acetyllactosamine (PL; (-3Gal␤1-4GlcNAc␤1-) n ) sequences, complex-type biantennary N-glycans, or novel chitin-derived glycans modified to contain terminal LN. Although terminal Gal residues are important for dGal-1 recognition, dGal-1 bound similarly to ␣3-sialylated and ␣2-fucosylated terminal LN, but not to ␣6-sialylated and ␣3-fucosylated terminal LN. The binding specificity of human recombinant dGal-1 was similar to that observed with purified bovine heart-derived dGal-1. Unexpectedly, dGal-1 bound free ligands in solution with relatively low affinity and displayed no preference for extended glycans, indicating that dGal-1 preferentially recognizes extended glycans only when they are surface-bound, such as found on cell surfaces. Human dGal-1 also bound to both native and desialylated human promyelocytic HL-60 cells with similar affinity as observed for immobilized long chain PL. Binding to these cells was reduced upon treatment with endo-␤-galactosidase, which cleaves PL sequences, indicating that cell-surface PLs are ligands. To test the role of dimerization in dGal-1 binding, we examined the binding of a mutated form of dGal-1 that weakly dimerizes (monomeric Gal-1 (mGal-1)) and a covalently dimerized (chemically cross-linked) form of mGal-1 (cd-mGal-1). dGal-1 and cd-mGal-1 had similar affinities that were both ϳ3.5-fold higher for immobilized PL than observed for mGal-1, suggesting that dGal-1 acts as a dimer to cross-link terminal LN units on immobilized PL. These results indicate that dGal-1 functions as a dimer to recognize LN units on extended PLs on cell surfaces.

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Vertebrate lectins, Comparison of properties of beta-galactoside-binding lectins from tissues of calf and chicken.

Cited by 131 publications

References 9 publications

Dissociation of the carbohydrate-binding and splicing activities of galectin-1

Dissociation of the carbohydrate-binding and splicing activities of galectin-1

Endogenous mammalian lectin localized extracellularly in lung elastic fibers

Dimeric Galectin-1 Binds with High Affinity to α2,3-Sialylated and Non-sialylated Terminal N-Acetyllactosamine Units on Surface-bound Extended Glycans

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