BIOMOLPEP group, Instituto de Biociências, UNESP, Campus do Litoral Paulista, São Vicente CEP 11380-972, São Paulo 11380-972, Brazil.
Departamento de Botânica, Instituto de Biociências, Universidade de São Paulo, São Paulo CEP 05508-090, São Paulo, Brazil.
Molecules. 2020 Jan 9;25(2):282. doi: 10.3390/molecules25020282.
Bioaffinity capturing of molecules allows the discovery of bioactive compounds and decreases the need for various stages in the natural compound isolation process. Despite the high selectivity of this technique, the screening and identification methodology depends on the presence of a protein to capture potential ligands. However, some proteins, such as snake secretory phospholipase A2 (sPLA2), have never been investigated using this approach. The purpose of this study was to evaluate the use of a new method for screening natural compounds using a bioaffinity-guided ultrafiltration method on sPLA2 followed by HPLC-MS to identify the compounds, and this method could be used to discover new anti-inflammatory compounds from the various organisms originating from biodiversity. Different extracts were selected to evaluate their ability to inhibit sPLA2 activity. The extracts were incubated with sPLA2 and the resulting mixture was ultrafiltrated to elute unbound components. The resulting compounds were identified by HPLC-MS. We identified hispidulin as one of the components present in the leaf and evaluated the ability of this isolated compound to neutralize the inflammatory activity of sPLA2 from .
生物亲和捕获分子可用于发现具有生物活性的化合物,并减少天然化合物分离过程中各个阶段的需求。尽管该技术具有高选择性,但筛选和鉴定方法取决于是否存在用于捕获潜在配体的蛋白质。然而,一些蛋白质,如蛇分泌型磷脂酶 A2(sPLA2),从未使用这种方法进行过研究。本研究旨在评估使用一种新方法,通过 sPLA2 上的亲和超滤筛选天然化合物,然后通过 HPLC-MS 鉴定化合物,这种方法可用于从生物多样性来源的各种生物体中发现新的抗炎化合物。选择了不同的提取物来评估它们抑制 sPLA2 活性的能力。将提取物与 sPLA2 一起孵育,然后将所得混合物超滤以洗脱未结合的成分。通过 HPLC-MS 鉴定所得化合物。我们鉴定出桦木脂素是存在于叶中的一种成分,并评估了这种分离化合物中和 的 sPLA2 炎症活性的能力。