期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 113, 期 5, 页码 1202-1207出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1519214113
关键词
synthetic biology; protein engineering; nanobiotechnology; split protein; antibody
资金
- Medical Research Council
- Merton College Oxford
- Sony
- Ecole Normale Superieure de Lyon
- Royal Society
- European Research Council [ERC-2013-CoG 615945-PeptidePadlock]
- Medical Research Council [G1000819, 1243569] Funding Source: researchfish
- MRC [G1000819] Funding Source: UKRI
Programmed connection of amino acids or nucleotides into chains introduced a revolution in control of biological function. Reacting proteins together is more complex because of the number of reactive groups and delicate stability. Here we achieved sequence-programmed irreversible connection of protein units, forming polyprotein teams by sequential amidation and transamidation. SpyTag peptide is engineered to spontaneously form an isopeptide bond with SpyCatcher protein. By engineering the adhesin RrgA from Streptococcus pneumoniae, we developed the peptide SnoopTag, which formed a spontaneous isopeptide bond to its protein partner SnoopCatcher with >99% yield and no cross-reaction to SpyTag/SpyCatcher. Solid-phase attachment followed by sequential SpyTag or SnoopTag reaction between building-blocks enabled iterative extension. Linear, branched, and combinatorial polyproteins were synthesized, identifying optimal combinations of ligands against death receptors and growth factor receptors for cancer cell death signal activation. This simple and modular route to programmable polyproteams should enable exploration of a new area of biological space.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据