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蛇毒磷脂酶 A 与 magic-sized 量子点的体外示踪

In vitro tracking of phospholipase A from snake venom conjugated with magic-sized quantum dots.

机构信息

Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Uberlândia, MG, Brazil.

Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Uberlândia, MG, Brazil; Instituto de Química, Universidade Estadual Paulista, Araraquara, SP, Brazil.

出版信息

Int J Biol Macromol. 2019 Feb 1;122:461-468. doi: 10.1016/j.ijbiomac.2018.10.185. Epub 2018 Oct 29.

Abstract

Phospholipases A represent a family of enzymes with important application in medicine. However, direct tracking is difficult due to the absence of a stable, effective and specific marker for these enzymes. Magic-sized quantum dots (MSQDs) are inorganic semiconducting nanocrystals with unique physical properties. They have the ability to conjugate to proteins, making them excellent markers for biological systems. In this work, we labelled phospholipase A from Bothrops alternatus snake venom with Cadmium selenide (CdSe)/cadmium sulphate (CdS) MSQDs-a biocompatible and luminescent probe-. Bioconjugation was confirmed using infrared spectra and fluorescence microscopy, which demonstrated that the CdSe/CdS MSQDs interact with phospholipase A without interfering with its activity. This probe may be an important tool for the elucidation of many biological mechanisms, because it allows the pathway of phospholipase A to be tracked from its entry through the plasma membrane until its incorporation into the nucleus of myoblasts.

摘要

磷脂酶 A 是一类具有重要医学应用价值的酶。然而,由于缺乏这些酶的稳定、有效和特异性标记物,直接跟踪这些酶非常困难。 魔角量子点(MSQDs)是具有独特物理性质的无机半导体纳米晶体。它们能够与蛋白质结合,使它们成为生物系统的优秀标记物。在这项工作中,我们用硫化镉(CdS)/硒化镉(CdSe)MSQDs 标记了来自矛头蝮蛇蛇毒的磷脂酶 A——一种生物相容性和发光探针。使用红外光谱和荧光显微镜证实了生物结合,表明 CdSe/CdS MSQDs 与磷脂酶 A 相互作用而不干扰其活性。这种探针可能是阐明许多生物学机制的重要工具,因为它可以追踪磷脂酶 A 的途径,从其穿过质膜进入到肌母细胞的核内。

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