Galectins and other endogenous carbohydrate-binding proteins of animal bladder.
Defects in the glycocalyx of the bladder epithelium may be related to the development of bladder diseases including interstitial cystitis which is a chronic bladder disease of unknown etiology. Indirect evidence has implicated alterations in the bladder epithelial glycoconjugates in interstitial cystitis and vesicaler instillation of glycosaminoglycans is promoted as treatments. However, information on the nature of the glycoconjugates of the bladder epithelium and lectins that may interact with the exogenous instilled glycoconjugates is very limited. We have examined the endogenous lectin associated with bladder epithelium by immunohistochemistry using biotinylated neoglycoconjugates. The strong calcium-independent binding of beta-D-galactose probe suggested the presence of galectins in rabbit and human bladder. Extracts of rabbit bladder organ cultures metabolically labeled with [14C]-amino acids were subjected to affinity chromatography on immobilized lactose and the specifically bound material eluted with 0.2 M lactose. SDS-PAGE of the recovered proteins revealed a major band of approximately 30 kDa and a minor band of 21 kDa. Polymerase chain reaction and northern blot analysis showed that both galectin-3 and galectin-4 are expressed in rabbit bladder. Since galectin-3 from rabbit had been previously cloned, we cloned and sequenced galectin-4 from rabbit bladder. The deduced full length sequence of 328 amino acids revealed four distinct regions: a N-terminal peptide of 19 residues, two carbohydrate recognition domains of 130 residues each, and a linker region of 49 residues. Comparison of the rabbit galectin-4 sequence with those of human, pig, rat, and mouse revealed two invariant peptide motifs that are proposed as signature sequences for identifying related galectins.
Bhavanandan
V P
VP
Department of Biochemistry & Molecular Biology, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, USA.
Puch
S
S
Guo
X
X
Jiang
W
W
D000818:Animals; D001483:Base Sequence; D050260:Carbohydrate Metabolism; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D037502:Galectin 3 / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D037541:Galectin 4 / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D037161:Galectins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006001:Glycoconjugates / Q000378:metabolism; D006651:Histocytochemistry; D006801:Humans; D066298:In Vitro Techniques; D037102:Lectins / Q000302:isolation & purification / Q000378:metabolism; D009092:Mucous Membrane / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011817:Rabbits; D014513:Ureter / Q000378:metabolism; D001743:Urinary Bladder / Q000378:metabolism*
D018076:DNA, Complementary; D037502:Galectin 3; D037541:Galectin 4; D037161:Galectins; D006001:Glycoconjugates; D037102:Lectins