Thermodynamic binding studies of lectins from the diocleinae subtribe to deoxy analogs of the core trimannoside of asparagine-linked oligosaccharides.
Lectins from seven different species of the Diocleinae subtribe have been recently isolated and characterized in terms of their carbohydrate binding specificities (Dam, T. K., Cavada, B. S., Grangeiro, T. B., Santos, C. F., de Sousa, F. A. M., Oscarson, S., and Brewer, C. F. (1998) J. Biol. Chem. 273, 12082-12088). The lectins included those from Canavalia brasiliensis, Cratylia floribunda, Dioclea rostrata, Dioclea virgata, Dioclea violacea, and Dioclea guianensis. All of the lectins exhibited specificity for Man and Glc residues, but much higher affinities for the branched chain trimannoside, 3,6-di-O-(alpha-d-mannopyranosyl)-d-mannose, which is found in the core region of all asparagine-linked carbohydrates. In the present study, isothermal titration microcalorimetry is used to determine the binding thermodynamics of the above lectins, including a new lectin from Canavalia grandiflora, to a complete series of monodeoxy analogs of the core trimannoside. From losses in the affinity constants and enthalpies of binding of certain deoxy analogs, assignments are made of the hydroxyl epitopes on the trimannoside that are involved in binding to the lectins. The pattern of binding of the deoxy analogs is similar for all seven lectins, and similar to that of concanavalin A which is also a member of the Diocleinae subtribe. However, differences in the magnitude of the thermodynamic binding parameters of the lectins are observed, even though the lectins possess conserved contact residues in many cases, and highly conserved primary sequences. The results indicate that non-contact residues in the lectins, even those distant from the binding sites, modulate their thermodynamic binding parameters.
Dam
T K
TK
Departments of Molecular Pharmacology and Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Cavada
B S
BS
Grangeiro
T B
TB
Santos
C F
CF
Ceccatto
V M
VM
de Sousa
F A
FA
Oscarson
S
S
Brewer
C F
CF
D000595:Amino Acid Sequence; D002151:Calorimetry; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D017124:Conserved Sequence; D037102:Lectins / Q000737:chemistry*; D008362:Mannosides / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry*; D010802:Phylogeny; D037121:Plant Lectins; D010944:Plants / Q000737:chemistry*; D011485:Protein Binding; D012210:Rhamnose / Q000031:analogs & derivatives; D013816:Thermodynamics; D014312:Trisaccharides / Q000737:chemistry
D037102:Lectins; D008362:Mannosides; D009844:Oligosaccharides; D037121:Plant Lectins; D014312:Trisaccharides; D012210:Rhamnose