title: Aqueous zymography screening of matrix metalloproteinase activity and inhibition based on colorimetric gold nanoparticles.
abstract: An optical gold nanoparticles (AuNPs)-based method was fabricated for the rapid detection of matrix metalloproteinase (MMP) activity and screening potential MMP inhibitors without sophisticated instruments. The diagnosis platform was composed of AuNPs, particular MMP substrates and 6-mercapto-1-hexanol (MCH). The functionalized AuNPs were subjected to specific MMP digestion, and the MMP found the substrate on AuNPs, such that the AuNPs lost shelter and MCH increased the attraction force between AuNPs. Consequently, AuNPs aggregation and a color change from red to purple with increasing MMP concentration were observed. The surface plasmon resonance (SPR) of the formed AuNPs allowed for the quantitative detection of MMP activity. A sensitive linear correlation existed between the absorbance and the activity of the MMPs, which ranged from 10 ng/mL to 700 ng/mL in NTTC buffer and plasma samples. The proposed colorimetric method could be accomplished in a homogeneous solution with one-step operation in 30 min and has been successfully applied to the determination of particular MMP activity in plasma samples, in which the results are consistent with substrate zymography. This technology may become a simple platform for parallel screening a number of inhibitors and offer an alternative method to studying the efficiency of inhibitors for suppressing MMP activity. The absorbance ratio at 625 nm and 525 nm (A(625)/A(525)) confirmed the efficiency of the inhibitors as observed in substrate zymography. The IC(50) of ONO-4817 and galardin for MMP-1, MMP-2 and MMP-7 determined by the proposed colorimetric method was similar to the results of substrate zymography.
authors.affiliation: Department of Biological Science and Technology, National Chiao Tung University, Hsinchu 30068, Taiwan.
authors.forename: Yao-Chen
authors.initials: YC
authors.lastname: Chuang
authors.forename: Wei-Ting
authors.initials: WT
authors.lastname: Huang
authors.forename: Pin-Hsuan
authors.initials: PH
authors.lastname: Chiang
authors.forename: Meng-Che
authors.initials: MC
authors.lastname: Tang
authors.forename: Chih-Sheng
authors.initials: CS
authors.lastname: Lin
mesh_terms: D003124:Colorimetry / Q000379:methods; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006046:Gold / Q000737:chemistry*; D000441:Hexanols / Q000378:metabolism; D006801:Humans; D008297:Male; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000097:blood* / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013438:Sulfhydryl Compounds / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: C503488:6-mercapto-1-hexanol; D004791:Enzyme Inhibitors; D000441:Hexanols; D061965:Matrix Metalloproteinase Inhibitors; D013438:Sulfhydryl Compounds; D006046:Gold; D020782:Matrix Metalloproteinases