Quantitative measurements of vancomycin binding to self-assembled peptide monolayers on chips by quartz crystal microbalance.
This paper describes direct binding of a small vancomycin to peptide ligands immobilized on a sensor chip using quartz crystal microbalance. In this study, the binding ligands were composed of three components: a molecular recognition element (peptide), a conformationally flexible and hydrophilic linker, and a long-chain alkanethiol. These peptide ligands were used to establish the well-packed, self-assembled monolayers on quartz chips and could be readily synthesized using conventional organic chemistry protocols. Results of quartz crystal microbalance measurements showed that vancomycin specifically associated with the d-Ala-d-Ala-containing peptide with an affinity of 3.2+/-0.3 microM and was, as expected, completely inactive to the self-assembled monolayer presenting l-Ala-l-Ala peptide. The dissociation constant obtained correlated well with values reported in literature and was further confirmed by surface plasmon resonance measurement (2.7+/-0.7 microM). The technique used in this study should be applicable to both peptidyl and nonpeptidyl ligands of greater complexity than that used here. This method is practical, it provides quantitative binding information, and complicated analysis is avoided.
Tseng
Ming-Chung
MC
Department of Chemistry and Biochemistry, National Chung Cheng University, Chia-Yi, Taiwan 621, Republic of China.
Chang
Yi-Pin
YP
Chu
Yen-Ho
YH
D000409:Alanine / Q000737:chemistry; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D003460:Crystallization; D004867:Equipment Design; D007700:Kinetics; D008024:Ligands; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011791:Quartz / Q000737:chemistry*; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance; D014640:Vancomycin / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D008024:Ligands; D010455:Peptides; D011791:Quartz; D014640:Vancomycin; D000409:Alanine