Novel human cell expression method reveals the role and prevalence of posttranslational modification in nonmuscle tropomyosins.

ABSTRACT: Biochemical studies require large quantities of proteins, which are typically obtained using bacterial overexpression. However, the folding machinery in bacteria is inadequate for expressing many mammalian proteins, which additionally undergo posttranslational modifications (PTMs) that bacteria, yeast, or insect cells cannot perform. Many proteins also require native N- and C-termini and cannot tolerate extra tag amino acids for proper function. Tropomyosin (Tpm), a coiled coil protein that decorates most actin filaments in cells, requires both native N- and C-termini and PTMs, specifically N-terminal acetylation (Nt-acetylation), to polymerize along actin filaments. Here, we describe a new method that combines native protein expression in human cells with an intein-based purification tag

SUBMITTER: Carman PJ 

PROVIDER: S-EPMC8463859 | biostudies-literature | 2021 Oct

REPOSITORIES: biostudies-literature