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Purification of Human β and γ Actin from Budding Yeast

ABSTRACT: To facilitate biochemical studies of human cytoplasmic actins, Saccharomyces cerevisiaestrains were developed that express either human β-actin or γ-actin as their sole actin. Actins purified from these strains have biochemical properties similar to mammalian skeletal muscle α-actin, but display differential binding to some key actin regulators. Mass-spectrometry analysis shows that the human actins made from yeast generally retain their N-terminal methionines, which are modified by acetylation.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human) Oryctolagus Sp. 'rabbit_od' Saccharomyces Cerevisiae (baker's Yeast)

TISSUE(S): Cell Culture, Femoral Muscle

SUBMITTER: Ebbing de Jong

LAB HEAD: Jessica Henty-Ridilla

PROVIDER: PXD040174 | Pride | 2023-04-24

REPOSITORIES: Pride

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Purification of human β- and γ-actin from budding yeast.

Haarer Brian K BK Pimm Morgan L ML de Jong Ebbing P EP Amberg David C DC Henty-Ridilla Jessica L JL

Journal of cell science 20230509 9

Biochemical studies of human actin and its binding partners rely heavily on abundant and easily purified α-actin from skeletal muscle. Therefore, muscle actin has been used to evaluate and determine the activities of most actin regulatory proteins but there is an underlying concern that these proteins perform differently from actin present in non-muscle cells. To provide easily accessible and relatively abundant sources of human β- or γ-actin (i.e. cytoplasmic actins), we developed Saccharomyces ...[more]

PMID: 37070275

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