@inbook{209c8b03cc0e4fdf8ddb8c1678f7bba5,
title = "Expression and purification of the type II metacaspase from a unicellular green alga Chlamydomonas reinhardtii",
abstract = "Type II metacaspases (MCAs) are proteases, belonging to the C14B MEROPS family. Like the MCAs of type I and type III, they preferentially cleave their substrates after the positively charged amino acid residues (Arg or Lys) at the P1 position. Type II MCAs from various higher plants have already been successfully overexpressed in E. coli mostly as His-tagged proteins and were shown to be proteolytically active after the purification. Here we present a protocol for expression and purification of the only type II MCA from the model green alga Chlamydomonas reinhardtii. The two-step purification, which consists of immobilized metal affinity chromatography using cobalt as ion followed by size-exclusion chromatography, can be performed in 1 day and yields 4 mg CrMCA-II protein per liter of overexpression culture.",
keywords = "Cobalt IMAC, Metacaspase, Overexpression, Protease, Cobalt IMAC, Metacaspase, Overexpression, Protease",
author = "Igor Sabljic and Yong Zou and Marina Klemen{\v c}i{\v c} and Christiane Funk and Jerry St{\aa}hlberg and Peter Bozhkov",
year = "2022",
doi = "10.1007/978-1-0716-2079-3\_2",
language = "English",
isbn = "978-1-0716-2078-6",
series = "Methods in Molecular Biology",
publisher = "Humana",
number = "2447",
pages = "13--20",
booktitle = "Plant Proteases and Plant Cell Death : Methods and Protocols",
}