ZG16p，一种植物β-晶状折叠凝集素的动物同源物：天然和重组 ZG16p 的纯化方法以及使用 ZG16p 抑制癌细胞生长的抑制测定。

ZG16p, an Animal Homologue of Plant β-Prism Fold Lectins: Purification Methods of Natural and Recombinant ZG16p and Inhibition Assay of Cancer Cell Growth Using ZG16p.

机构信息

Natural Science Division, Faculty of Science, Ochanomizu University, Tokyo, Japan.

Natural Science Division, Faculty of Core Research, Ochanomizu University, Tokyo, Japan.

出版信息

Methods Mol Biol. 2020;2132:339-347. doi: 10.1007/978-1-0716-0430-4_33.

DOI:10.1007/978-1-0716-0430-4_33

PMID:32306341

Abstract

ZG16p is a soluble 16-kDa protein abundantly expressed in the pancreas and gut, and has a β-prism fold structure similar to that of mannose-binding Jacalin-related lectins (mJRLs) such as BanLec, Heltuba, and Artocarpin. ZG16p binds to mannose via the well-conserved GXXXD loop among mJRLs and sulfated glycosaminoglycans (e.g., heparin and heparan sulfate) via the basic patch of molecular surface. In addition to the above binding activities, ZG16p has inhibitory activity against proliferation of colon cancer cells. This manuscript describes purification of rat pancreatic ZG16p and recombinant ZG16p expressed in Escherichia coli expression system, and cell growth inhibition assay using ZG16p as an inhibitor.

摘要

ZG16p 是一种可溶性的 16kDa 蛋白，在胰腺和肠道中大量表达，其 β-棱柱折叠结构与甘露糖结合 Jacalin 相关凝集素（mJRL）相似，如 BanLec、Heltuba 和 Artocarpin。ZG16p 通过 mJRL 中保守的 GXXXD 环结合甘露糖，并通过分子表面的碱性斑块结合硫酸化糖胺聚糖（如肝素和硫酸乙酰肝素）。除了上述结合活性外，ZG16p 还具有抑制结肠癌细胞增殖的活性。本文描述了大鼠胰腺 ZG16p 的纯化和在大肠杆菌表达系统中表达的重组 ZG16p，以及使用 ZG16p 作为抑制剂的细胞生长抑制试验。