Difference in the binding mode of two mannose-binding proteins: demonstration of a selective minicluster effect.
Serum-type and liver-type mannose-binding proteins (MBP) are both present in higher animals and both are composed of a carbohydrate-recognition domain (CRD) and a collagenous domain. Although known as mannose-binding proteins, these proteins bind N-acetylglucosamine and other related sugars quite well. An earlier specificity study using cloned CRD portions of both types of MBP from rat [Childs, R. A., Feizi, T., Yuen, C.-T., Drickamer, K., & Quesenberry, M. (1990) J. Biol. Chem. 265, 20770-20777] revealed that the liver MBP CRD binds the trimannosyl core structure of N-glycosides, whereas the serum MBP CRD does not. We studied the substrate preferences of these CRDs using both solid and solution phase assays, testing monosaccharides, glycoproteins, and synthetic cluster ligands. While there was no significant difference in the monosaccharide binding specificities of the two CRDs, they displayed very different affinities for natural glycoproteins and mannose-containing cluster glycosides. Most interestingly, synthetic cluster ligands with two terminal GlcNAc moieties have affinity equal to monovalent GlcNAc ligands toward both CRDs, whereas a series of structurally similar Man-terminated divalent ligand displays about 20-fold enhanced affinity toward liver CRD only. A plausible explanation is that the liver MBP CRD has two sugar binding sites per subunit, one of which binds only mannose, and the other, both mannose and N-acetylglucosamine. In contrast, the serum MBP CRD has only one site of the latter type. Results of isothermal titration calorimetry support this hypothesis.
Quesenberry
M S
MS
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218, USA.
Lee
R T
RT
Lee
Y C
YC
D001665:Binding Sites; D002151:Calorimetry; D002240:Carbohydrate Sequence; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D006023:Glycoproteins / Q000494:pharmacology; D006027:Glycosides / Q000737:chemistry; D008358:Mannose / Q000737:chemistry*; D037241:Mannose-Binding Lectins; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000737:chemistry; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D017186:Titrimetry
D002352:Carrier Proteins; D006023:Glycoproteins; D006027:Glycosides; D037241:Mannose-Binding Lectins; D009005:Monosaccharides; D008358:Mannose