Abstract
In Saccharomyces cerevisiae, the membrane-associated enzyme phosphatidylserine synthase (EC 2.7.8.8) is present in the mitochondria and the endoplasmic reticulum. The enzyme from both membrane fractions reacted with antiserum raised against a hybrid protein expressed from a TRPE-CHO1 fusion gene in Escherichia coli and was absent in a cho1 null mutant, strongly suggesting that both the mitochondrial and microsomal forms of phosphatidylserine synthase are the products of the CHO1 gene. The highest degree of purification of enzymatically active protein was 380- and 420-fold from the mitochondrial and the microsomal compartments, respectively. In both cases, the enzymatically active and immunoreactive material comigrated with a protein band of 30,000 apparent molecular weight. In the absence of protease inhibitors during the preparation of membranes, the enzyme underwent degradation to an enzymatically active protein of 23,000 apparent molecular weight.
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- Atkinson K., Fogel S., Henry S. A. Yeast mutant defective in phosphatidylserine synthesis. J Biol Chem. 1980 Jul 25;255(14):6653–6661. [PubMed] [Google Scholar]
- Bae-Lee M. S., Carman G. M. Phosphatidylserine synthesis in Saccharomyces cerevisiae. Purification and characterization of membrane-associated phosphatidylserine synthase. J Biol Chem. 1984 Sep 10;259(17):10857–10862. [PubMed] [Google Scholar]
- Bailis A. M., Poole M. A., Carman G. M., Henry S. A. The membrane-associated enzyme phosphatidylserine synthase is regulated at the level of mRNA abundance. Mol Cell Biol. 1987 Jan;7(1):167–176. doi: 10.1128/mcb.7.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carlson M., Botstein D. Two differentially regulated mRNAs with different 5' ends encode secreted with intracellular forms of yeast invertase. Cell. 1982 Jan;28(1):145–154. doi: 10.1016/0092-8674(82)90384-1. [DOI] [PubMed] [Google Scholar]
- Kiyono K., Miura K., Kushima Y., Hikiji T., Fukushima M., Shibuya I., Ohta A. Primary structure and product characterization of the Saccharomyces cerevisiae CHO1 gene that encodes phosphatidylserine synthase. J Biochem. 1987 Nov;102(5):1089–1100. doi: 10.1093/oxfordjournals.jbchem.a122147. [DOI] [PubMed] [Google Scholar]
- Kuchler K., Daum G., Paltauf F. Subcellular and submitochondrial localization of phospholipid-synthesizing enzymes in Saccharomyces cerevisiae. J Bacteriol. 1986 Mar;165(3):901–910. doi: 10.1128/jb.165.3.901-910.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Letts V. A., Klig L. S., Bae-Lee M., Carman G. M., Henry S. A. Isolation of the yeast structural gene for the membrane-associated enzyme phosphatidylserine synthase. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7279–7283. doi: 10.1073/pnas.80.23.7279. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poole M. A., Fischl A. S., Carman G. M. Enzymatic detection of phospholipid biosynthetic enzymes after electroblotting. J Bacteriol. 1985 Feb;161(2):772–774. doi: 10.1128/jb.161.2.772-774.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poole M. A., Homann M. J., Bae-Lee M. S., Carman G. M. Regulation of phosphatidylserine synthase from Saccharomyces cerevisiae by phospholipid precursors. J Bacteriol. 1986 Nov;168(2):668–672. doi: 10.1128/jb.168.2.668-672.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]