A fluorescent lectin array using supramolecular hydrogel for simple detection and pattern profiling for various glycoconjugates

被引:121
作者
Koshi, Yoichiro [1 ]
Nakata, Eiji [1 ]
Yamane, Hiroki [1 ]
Hamachi, Itaru [1 ]
机构
[1] Kyoto Univ, PRESTO, Dept Synthet Chem & Biol Chem, JST,Nishikyo Ku, Kyoto 6158510, Japan
关键词
D O I
10.1021/ja0613963
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Because sugar and its derivatives play important roles in various biological phenomena, the rapid and high-throughput analysis of various glycoconjugates is keenly desirable. We describe herein the construction of a novel fluorescent lectin array for saccharide detection using a supramolecular hydrogel matrix. In this array, the fluorescent lectins were noncovalently fixed under semi-wet conditions to suppress the protein denaturation. It is demonstrated by fluorescence titration and fluorescence lifetime experiments that the immobilized lectins act as a molecular recognition scaffold in the hydrogel matrix, similar to that in aqueous solution. That is, a bimolecular fluorescence quenching and recovery (BFQR) method can successfully operate under both conditions. This enables one to fluorescently read-out a series of saccharides on the basis of the recognition selectivity and affinity of the immobilized lectins without tedious washing processes and without labeling the target saccharides. Simple and high-throughput sensing and profiling were carried out using the present lectin array for diverse glycoconjugates, which not only included a simple glucose, but also oligosaccharides, and glycoproteins, and, furthermore, the pattern recognition and profiling of several types of cell lysates were also accomplished.
引用
收藏
页码:10413 / 10422
页数:10
相关论文
共 62 条
[11]   BANDEIRAEA-SIMPLICIFOLIA LECTIN DEMONSTRATES SIGNIFICANTLY MORE CAPILLARIES IN RAT SKELETAL-MUSCLE THAN ENZYME METHODS [J].
CHRISTIE, KN ;
THOMSON, C .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1989, 37 (08) :1303-1304
[12]   Thermodynamic studies of lectin-carbohydrate interactions by isothermal titration calorimetry [J].
Dam, TK ;
Brewer, CF .
CHEMICAL REVIEWS, 2002, 102 (02) :387-429
[13]   The use of carbohydrate microarrays to study carbohydrate-cell interactions and to detect pathogens [J].
Disney, MD ;
Seeberger, PH .
CHEMISTRY & BIOLOGY, 2004, 11 (12) :1701-1707
[14]   Targeting glycosylation as a therapeutic approach [J].
Dwek, RA ;
Butters, TD ;
Platt, FM ;
Zitzmann, N .
NATURE REVIEWS DRUG DISCOVERY, 2002, 1 (01) :65-75
[15]   Glycobiology: Toward understanding the function of sugars [J].
Dwek, RA .
CHEMICAL REVIEWS, 1996, 96 (02) :683-720
[16]   Application of lectin microarray to crude samples: Differential glycan profiling of Lec mutants [J].
Ebe, Y ;
Kuno, A ;
Uchiyama, N ;
Koseki-Kuno, S ;
Yamada, M ;
Sato, T ;
Narimatsu, H ;
Hirabayashi, J .
JOURNAL OF BIOCHEMISTRY, 2006, 139 (03) :323-327
[17]   Water gelation by small organic molecules [J].
Estroff, LA ;
Hamilton, AD .
CHEMICAL REVIEWS, 2004, 104 (03) :1201-1217
[18]  
GREEN ED, 1988, J BIOL CHEM, V263, P18253
[19]   Solid phase lipid synthesis (SPLS) for construction of an artificial glycolipid library [J].
Hamachi, I ;
Kiyonaka, S ;
Shinkai, S .
CHEMICAL COMMUNICATIONS, 2000, (14) :1281-1282
[20]   Composition of N-linked carbohydrates from ovalbumin and Co-purified glycoproteins [J].
Harvey, DJ ;
Wing, DR ;
Küster, B ;
Wilson, IBH .
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY, 2000, 11 (06) :564-571