荧光检测 MMP-9。I. MMP-9 特异性地裂解 Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys 肽。
Fluorescence detection of MMP-9. I. MMP-9 selectively cleaves Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys peptide.
机构信息
Center of Commercialization of Fluorescence Technologies, Department of Molecular Biology & Immunology, UNTHSC, Fort Worth, TX 76107, USA.
出版信息
Curr Pharm Biotechnol. 2011 May;12(5):834-8. doi: 10.2174/138920111795470967.
MMP-9 enzyme recognizes a peptide sequence Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys and cleaves the peptide into two parts. We synthesized a dual fluorophore beacon consisting of 5-FAM and Cy5 dyes. The fluorescence emission of the fluorescein moiety is dramatically quenched by Cy5 molecule due to Förster Resonance Energy Transfer (FRET) and the fluorescence of Cy5 is strongly enhanced. Upon addition of MMP-9 enzyme, the fluorescence of 5-FAM intensifies and Cy5 decreases. The control MMP-2 enzyme does not cause any changes in either 5-FAM or Cy5 fluorescence. We believe that our observation will help in early detection of elevated MMP-9 levels under disease conditions.
MMP-9 酶识别肽序列 Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys，并将肽切割成两部分。我们合成了一个由 5-FAM 和 Cy5 染料组成的双荧光探针。由于Förster 共振能量转移（FRET），荧光素部分的荧光发射被 Cy5 分子显著猝灭，并且 Cy5 的荧光强烈增强。加入 MMP-9 酶后，5-FAM 的荧光增强，而 Cy5 的荧光减弱。对照 MMP-2 酶不会引起 5-FAM 或 Cy5 荧光的任何变化。我们相信我们的观察结果将有助于在疾病状态下早期检测升高的 MMP-9 水平。
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