Electroporated recombinant proteins as tools for in vivo functional complementation, imaging and chemical biology.

ABSTRACT: Delivery of native or chemically modified recombinant proteins into mammalian cells shows promise for functional investigations and various technological applications, but concerns that sub-cellular localization and functional integrity of delivered proteins may be affected remain high. Here, we surveyed batch electroporation as a delivery tool for single polypeptides and multi-subunit protein assemblies of the kinetochore, a spatially confined and well-studied subcellular structure. After electroporation into human cells, recombinant fluorescent Ndc80 and Mis12 multi-subunit complexes exhibited native localization, physically interacted with endogenous binding partners, and functionally complemented depleted endogenous counterparts to promote mitotic checkpoint signaling and chromosome se

SUBMITTER: Alex A 

PROVIDER: S-EPMC6656429 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature