The Ca(2+)-dependent carbohydrate-recognition domain (CRD) of rat serum mannose-binding protein has been subjected to site-directed mutagenesis to determine the importance of individual residues in ligation of mannose and related sugars. The effects of the mutations were assessed by direct binding assays, competition binding studies, partial proteolysis, and NMR analysis of sugar-CRD titrations. As suggested by the crystal structure of the mannose-binding CRD complexed with oligosaccharide ligand, asparagine and glutamic acid residues that interact with hydroxyl groups 3 and 4 of the sugar, as well as with one of the two bound Ca2+, are critical for ligand binding. In addition, the beta-carbon of His189 contributes substantially to the binding affinity, apparently through a van der Waals contact with C-4 of the sugar ligand. van der Waals contacts between the imidazole ring of His189 and the 2 hydroxyl group of mannose, and between Ile207 and C-6 of mannose, observed in the crystal structure, contribute less to stability of the ligand complex. The effects of changes at positions 189 and 207 on the ability of the CRD to distinguish between alpha-and beta-methyl L-fucosides suggest that fucose may bind in an alternative orientation compared to the arrangement originally proposed based on the mannose-CRD complex.
[NMR paper] Analysis of competitive binding of ligands to human serum albumin using NMR relaxatio
Analysis of competitive binding of ligands to human serum albumin using NMR relaxation measurements.
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J Pharm Biomed Anal. 2004 Feb 4;34(2):247-54
Authors: Cui YF, Bai GY, Li CG, Ye CH, Liu ML
The competitive binding of two ligands, ibuprofen (IBP) and salicylic acid (SAL), to human serum albumin (HSA) was studied by using nuclear magnetic resonance (NMR) relaxation measurements. When the concentration of one ligand was...
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[NMR paper] Selective NMR observation of inhibitor and sugar binding to the galactose-H(+) sympor
Selective NMR observation of inhibitor and sugar binding to the galactose-H(+) symport protein GalP, of Escherichia coli.
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Biochim Biophys Acta. 2000 Dec 20;1509(1-2):55-64
Authors: Appleyard AN, Herbert RB, Henderson PJ, Watts A, Spooner PJ
The binding of the transport inhibitor forskolin, synthetically labelled with (13)C, to the galactose-H(+) symport protein GalP, overexpressed in its native inner...
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Defining a Stem Length-Dependent Binding Mechanism for the Cocaine-Binding Aptamer. A
Defining a Stem Length-Dependent Binding Mechanism for the Cocaine-Binding Aptamer. A Combined NMR and Calorimetry Study
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Biochemistry
DOI: 10.1021/bi100952k
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[NMR paper] H NMR study of the solution structure of Ac-AMP2, a sugar binding antimicrobial prote
H NMR study of the solution structure of Ac-AMP2, a sugar binding antimicrobial protein isolated from Amaranthus caudatus.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles H NMR study of the solution structure of Ac-AMP2, a sugar binding antimicrobial protein isolated from Amaranthus caudatus.
J Mol Biol. 1996 May 3;258(2):322-33
Authors: Martins JC, Maes D, Loris R, Pepermans HA, Wyns L, Willem R, Verheyden P
The conformation in water of antimicrobial protein 2 from...
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[NMR paper] Binding of sugar ligands to Ca(2+)-dependent animal lectins. I. Analysis of mannose b
Binding of sugar ligands to Ca(2+)-dependent animal lectins. I. Analysis of mannose binding by site-directed mutagenesis and NMR.
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J Biol Chem. 1994 Jun 3;269(22):15505-11
Authors: Iobst ST, Wormald MR, Weis WI, Dwek RA, Drickamer K
The Ca(2+)-dependent carbohydrate-recognition domain (CRD) of rat serum mannose-binding protein has been subjected to site-directed mutagenesis to determine the...
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[NMR paper] NMR observation of substrate in the binding site of an active sugar-H+ symport protei
NMR observation of substrate in the binding site of an active sugar-H+ symport protein in native membranes.
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Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3877-81
Authors: Spooner PJ, Rutherford NG, Watts A, Henderson PJ
NMR methods have been adopted to observe directly the characteristics of substrate...
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[NMR paper] NMR observation of substrate in the binding site of an active sugar-H+ symport protei
NMR observation of substrate in the binding site of an active sugar-H+ symport protein in native membranes.
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Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3877-81
Authors: Spooner PJ, Rutherford NG, Watts A, Henderson PJ
NMR methods have been adopted to observe directly the characteristics of substrate...
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[NMR paper] 19F NMR studies of the D-galactose chemosensory receptor. 1. Sugar binding yields a g
19F NMR studies of the D-galactose chemosensory receptor. 1. Sugar binding yields a global structural change.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc-MS.gif Related Articles 19F NMR studies of the D-galactose chemosensory receptor. 1. Sugar binding yields a global structural change.
Biochemistry. 1991 Apr 30;30(17):4248-56
Authors: Luck LA, Falke JJ
The Escherichia coli D-galactose and D-glucose receptor is an aqueous sugar-binding protein and the first component in the...