在 PVA 基质中寡肽连接给体-受体体系的 FRET 研究。

FRET study in oligopeptide-linked donor-acceptor system in PVA matrix.

机构信息

Institute of Molecular Medicine, Center for Fluorescence Technologies and Nanomedicine, University of North Texas Health Science Center, Fort Worth, TX 76107, USA.

出版信息

Methods Appl Fluoresc. 2016 Dec 13;4(4):047002. doi: 10.1088/2050-6120/4/4/047002.

DOI:10.1088/2050-6120/4/4/047002

PMID:28192309

Abstract

An oligopeptide: Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys-NH, cleaved specifically by a matrix metalloproteinase 9 (MMP-9) at the Ser-Leu bond, was labeled on the ε-NH groups of lysine with donor (5, 6 TAMRA) and acceptor (HiLyte647) dye. The donor control was a peptide labeled with 5, 6 TAMRA only on the C-terminal lysine, and the acceptor control was free HiLyte647. Following three products were studied by dissolving in 10% (w/w) poly(vinyl alcohol) and dried on glass slides forming 200 micron films. Absorption spectra of the films show full additivity of donor and acceptor absorptions. A strong Fluorescence Resonance Energy Transfer (FRET) with an efficiency of about 85% was observed in the fluorescence emission and excitation spectra. The lifetime of the donor was shorter and heterogeneous compared with the donor control.

摘要

一种寡肽

Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys-NH，在 Ser-Leu 键处被基质金属蛋白酶 9 (MMP-9) 特异性切割，其赖氨酸的 ε-NH 基团上标记了供体（5,6 TAMRA）和受体（HiLyte647）染料。供体对照肽仅在 C 末端赖氨酸上标记有 5,6 TAMRA，受体对照物为游离的 HiLyte647。将以下三种产物溶解在 10%（w/w）聚乙烯醇中并在玻璃载玻片上干燥形成 200 微米薄膜。薄膜的吸收光谱显示供体和受体吸收的完全加和性。在荧光发射和激发光谱中观察到强的荧光共振能量转移（FRET），效率约为 85%。与供体对照物相比，供体的寿命更短且不均匀。