SPR studies of carbohydrate-protein interactions: Signal enhancement of low-molecular-mass analytes by organoplatinum(II)-labeling

被引:55
作者
Beccati, D
Halkes, KM
Batema, GD
Guillena, G
de Souza, AC
van Koten, G
Kamerling, JP
机构
[1] Univ Utrecht, Dept Bioorgan Chem, Bijvoet Ctr, NL-3584 CH Utrecht, Netherlands
[2] Univ Utrecht, Debye Inst, Dept Met Mediated Synth, NL-3584 CH Utrecht, Netherlands
关键词
carbohydrates; lectin; organoplatinum(II) labeling; signal enhancement; surface plasmon resonance spectroscopy;
D O I
10.1002/cbic.200400402
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The relatively insensitive surface plosmon resonance (SPR) signal detection of low-molecular-moss onolytes that bind with weak affinity to a protein-for example, carbohydrate-lectin binding is hampering the use of biosensors in interaction studies. In this investigation, low-molecular-mass carbohydrates have been labeled with an organoplatinum(II) complex of the type [PtCl-(NCN-R)]. The attachment of this complex increased the SPR response tremendously and allowed the detection of binding events between monosaccharides and lectins at very low analyte concentrations. The platinum atom inside the orgonoplatinum(II) complex was shown to be essential for the SPR-signal enhancement. The organoplatinum(II) complex did not influence the specificity of the biological interaction, but both the signal enhancement and the different binding character of labeled compounds when compared with unlabeled ones makes the method unsuitable for the direct calculation of biologically relevant kinetic parameters. However, the labeling procedure is expected to be of high relevance for qualitative binding studies and relative affinity ranking of small molecules (not restricted only to carbohydrates) to receptors, a process of immense interest in pharmaceutical research.
引用
收藏
页码:1196 / 1203
页数:8
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