Departments of Molecular Medicine and Integrative Structural and Computational Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Departments of Molecular Medicine and Integrative Structural and Computational Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Bioorg Med Chem Lett. 2019 Sep 15;29(18):2609-2612. doi: 10.1016/j.bmcl.2019.07.054. Epub 2019 Aug 1.
To identify sialic acid binding proteins from complex proteomes, three photocrosslinking affinity-based probes were constructed using Neu5Ac (5 and 6) and Neu5Ac2en (7) scaffolds. Kinetic inhibition assays and Western blotting revealed the Neu5Ac2en-based 7 to be an effective probe for the labeling of a purified gut microbial sialidase (BDI_2946) and a purified human sialic acid binding protein (hCD33). Additionally, LC-MS/MS affinity-based protein profiling verified the ability of 7 to enrich a low-abundance sialic acid binding protein (complement factor H) from human serum thus validating the utility of this probe in a complex context.
为了从复杂的蛋白质组中鉴定唾液酸结合蛋白，我们构建了三个基于光交联亲和力的探针，使用 Neu5Ac（5 和 6）和 Neu5Ac2en（7）支架。动力学抑制测定和 Western blot 显示，基于 Neu5Ac2en 的 7 是一种有效的探针，可用于标记纯化的肠道微生物唾液酸酶（BDI_2946）和纯化的人唾液酸结合蛋白（hCD33）。此外，LC-MS/MS 基于亲和力的蛋白质谱分析验证了 7 从人血清中富集低丰度唾液酸结合蛋白（补体因子 H）的能力，从而验证了该探针在复杂环境中的实用性。
