Ligand Specificity of CS-35, a Monoclonal Antibody That Recognizes Mycobacterial Lipoarabinomannan: A Model System for Oligofuranoside-Protein Recognition

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• 作者：Christoph Rademacher ; Glen K. Shoemaker ; Hyo-Sun Kim ; Ruixiang Blake Zheng ; Hashem Taha ; Chunjuan Liu ; Ruel C. Nacario ; David C. Schriemer ; John S. Klassen ; Thomas Peters ; Todd L. Lowary
• 刊名：Journal of the American Chemical Society
• 出版年：2007
• 出版时间：August 29, 2007
• 年：2007
• 卷：129
• 期：34
• 页码：10489 - 10502
• 全文大小：357K
• 年卷期：v.129,no.34(August 29, 2007)
• ISSN：1520-5126

文摘

The CS-35 antibody is widely used in the characterization of glycans containing D-arabinofuranoseresidues, in particular polysaccharides present in the mycobacterial cell wall. A detailed understanding ofthe combining site of this antibody and the measurement of its binding to different ligands is of interest asthis knowledge will have implications in the characterization of arabinofuranose-containing glycoconjugatesthat are increasingly recognized as important biological molecules. Of even greater significance is that anin-depth study of this carbohydrate-protein interaction will provide insights into the mechanisms by whicholigosaccharides containing furanose rings are bound by proteins, an area that has, to date, received littleattention. This system has been refractory to X-ray crystallography, and thus we report here a study of theinteraction of CS-35 with its ligands using a combination of chemical synthesis, mass spectrometry, titrationmicrocalorimetry, and NMR spectroscopy. Through these investigations we have established that the bindingpocket recognizes, as a minimum epitope, a linear tetrasaccharide motif and that the residues at the reducingand non-reducing end of the oligosaccharide are essential for tight binding. The residue at the non-reducingend appears to be bound in an aliphatic pocket, whereas the rest of the tetrasaccharide interacts morestrongly with aromatic amino acids.