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Purification and some properties of Saccharomyces cerevisiae meiosis-specific protein kinase Ime2.
- Source :
-
Protein expression and purification [Protein Expr Purif] 2002 Dec; Vol. 26 (3), pp. 416-24. - Publication Year :
- 2002
-
Abstract
- Ime2 is the founding member of a family of protein kinases that are required for effective progression through meiotic development. Ime2 is essential for the induction of meiosis-specific genes and for the activation of meiotic DNA replication in the budding yeast Saccharomyces cerevisiae. Aside from the fact that Ime2 is a protein kinase and shares several amino acid motifs with cyclin dependent kinases, virtually nothing is known about its enzymatic properties or substrates. Biochemical characterization of Ime2 has been hindered by its low abundance and short half-life. We have created baculovirus expression vectors to produce recombinant Ime2 in insect cells. In this report, we describe the overproduction of Ime2 and its purification using affinity chromatography. Using this procedure, we have been able to purify up to 2mg Ime2 from 1L of infected insect cells. The Ime2 isolated by this method displays properties similar to those of the native enzyme that has been immunoprecipitated from yeast. The high level expression of Ime2 in this system and its ease of purification will be beneficial for more extensive biochemical analysis of Ime2 and related meiosis-specific kinases.
- Subjects :
- Animals
Cell Cycle Proteins genetics
Cell Line
Chromatography, Affinity
Guanosine Triphosphate metabolism
Histones metabolism
Intracellular Signaling Peptides and Proteins
Kinetics
Phosphorylation
Protein Kinases genetics
Protein Serine-Threonine Kinases
Recombinant Proteins genetics
Recombinant Proteins isolation & purification
Recombinant Proteins metabolism
Saccharomyces cerevisiae cytology
Saccharomyces cerevisiae genetics
Saccharomyces cerevisiae Proteins genetics
Spodoptera
Cell Cycle Proteins isolation & purification
Cell Cycle Proteins metabolism
Meiosis
Protein Kinases isolation & purification
Protein Kinases metabolism
Saccharomyces cerevisiae enzymology
Saccharomyces cerevisiae Proteins isolation & purification
Saccharomyces cerevisiae Proteins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1046-5928
- Volume :
- 26
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Protein expression and purification
- Publication Type :
- Academic Journal
- Accession number :
- 12460765
- Full Text :
- https://doi.org/10.1016/s1046-5928(02)00548-x