Crosslinking of Cys-mutated human galectin-1 to the model glycoprotein ligands asialofetuin and laminin by using a photoactivatable bifunctional reagent.
Galectins are a group of animal lectins characterized by their specificity for β-galactosides. In our previous study, we showed that a human galectin-1 (hGal-1) mutant, in which a cysteine residue was introduced at Lys(28), forms a covalently cross-linked complex with the model glycoprotein ligands asialofetuin and laminin by using the photoactivatable sulfhydryl reagent benzophenone-4-maleimide (BPM). In the present study, we used several hGal-1 mutants in which single cysteine residues were introduced at different positions and examined their ability to form a covalent complex with asialofetuin or laminin by using BPM. We found that the efficiency of formation of the cross-linked products differed depending on the positions of the cysteine introduced and also on the ligand used for crosslinking. Therefore, by using different cysteine hGal-1 mutants, the chances of isolating different ligands for hGal-1 should increase depending on the systems and cells used.
Faculty of Pharmaceutical Sciences, Josai University.
Mayumi
M
Tamura
Tomoe
T
Watanabe
Takanori
T
Igarashi
Tomoharu
T
Takeuchi
Ken-ichi
K
Kasai
Yoichiro
Y
Arata
D001212:Asialoglycoproteins / Q000737:chemistry*; D001577:Benzophenones / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry*; D003545:Cysteine / Q000737:chemistry* / Q000235:genetics; D060748:Fetuins / Q000737:chemistry*; D037483:Galectin 1 / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D007797:Laminin / Q000737:chemistry*; D008024:Ligands; D008301:Maleimides / Q000737:chemistry*; D016297:Mutagenesis, Site-Directed; D019920:Photoaffinity Labels / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry
D001212:Asialoglycoproteins; D001577:Benzophenones; D003432:Cross-Linking Reagents; D060748:Fetuins; D037483:Galectin 1; C445499:LGALS1 protein, human; D007797:Laminin; D008024:Ligands; D008301:Maleimides; D019920:Photoaffinity Labels; D011994:Recombinant Proteins; C017029:asialofetuin; C046109:benzophenone-4-maleimide; D003545:Cysteine