Abstract
We have previously shown that a Chinese hamster ovary (CHO) cell mutant defective in phosphatidylserine synthase I recovers the enzyme activity on transfection with a pssA cDNA clone isolated from the parental CHO-K1. The resultant transfectant, CDT-1, exhibited about 20-fold higher specific activity of the enzyme in the membrane fraction than CHO-K1 cells. Polyclonal antibodies against two peptides of the predicted pssA product cross-reacted with a membrane protein having an apparent molecular mass of 42 kDa, which was overproduced in CDT-1 cells. By immunoprecipitation with the antibody, phosphatidylserine synthase I activity as web as the 42-kDa protein was eliminated from solubilized membrane proteins of CDT-1 cells. Both the enzyme activity and the 42-kDa protein of CHO-K1 cells were enriched in the mitochondria-associated membrane fraction and the microsome fraction, but neither was enriched in the mitochondria fraction or the cytosol fraction. These results suggest that the pssA gene encodes phosphatidylserine synthase I.
| Original language | English |
|---|---|
| Pages (from-to) | 262-266 |
| Number of pages | 5 |
| Journal | FEBS Letters |
| Volume | 395 |
| Issue number | 2-3 |
| DOIs | |
| Publication status | Published - Oct 21 1996 |
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All Science Journal Classification (ASJC) codes
- Biophysics
- Structural Biology
- Biochemistry
- Molecular Biology
- Genetics
- Cell Biology
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Immunochemical identification of the pssA gene product as phosphatidylserine synthase I of Chinese hamster ovary cells. / Saito, Kyoko; Kuge, Osamu; Akamatsu, Yuzuru; Nishijima, Masahiro.
In: FEBS Letters, Vol. 395, No. 2-3, 21.10.1996, p. 262-266.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Immunochemical identification of the pssA gene product as phosphatidylserine synthase I of Chinese hamster ovary cells
AU - Saito, Kyoko
AU - Kuge, Osamu
AU - Akamatsu, Yuzuru
AU - Nishijima, Masahiro
PY - 1996/10/21
Y1 - 1996/10/21
N2 - We have previously shown that a Chinese hamster ovary (CHO) cell mutant defective in phosphatidylserine synthase I recovers the enzyme activity on transfection with a pssA cDNA clone isolated from the parental CHO-K1. The resultant transfectant, CDT-1, exhibited about 20-fold higher specific activity of the enzyme in the membrane fraction than CHO-K1 cells. Polyclonal antibodies against two peptides of the predicted pssA product cross-reacted with a membrane protein having an apparent molecular mass of 42 kDa, which was overproduced in CDT-1 cells. By immunoprecipitation with the antibody, phosphatidylserine synthase I activity as web as the 42-kDa protein was eliminated from solubilized membrane proteins of CDT-1 cells. Both the enzyme activity and the 42-kDa protein of CHO-K1 cells were enriched in the mitochondria-associated membrane fraction and the microsome fraction, but neither was enriched in the mitochondria fraction or the cytosol fraction. These results suggest that the pssA gene encodes phosphatidylserine synthase I.
AB - We have previously shown that a Chinese hamster ovary (CHO) cell mutant defective in phosphatidylserine synthase I recovers the enzyme activity on transfection with a pssA cDNA clone isolated from the parental CHO-K1. The resultant transfectant, CDT-1, exhibited about 20-fold higher specific activity of the enzyme in the membrane fraction than CHO-K1 cells. Polyclonal antibodies against two peptides of the predicted pssA product cross-reacted with a membrane protein having an apparent molecular mass of 42 kDa, which was overproduced in CDT-1 cells. By immunoprecipitation with the antibody, phosphatidylserine synthase I activity as web as the 42-kDa protein was eliminated from solubilized membrane proteins of CDT-1 cells. Both the enzyme activity and the 42-kDa protein of CHO-K1 cells were enriched in the mitochondria-associated membrane fraction and the microsome fraction, but neither was enriched in the mitochondria fraction or the cytosol fraction. These results suggest that the pssA gene encodes phosphatidylserine synthase I.
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UR - http://www.scopus.com/inward/citedby.url?scp=0030597170&partnerID=8YFLogxK
U2 - 10.1016/0014-5793(96)01049-6
DO - 10.1016/0014-5793(96)01049-6
M3 - Article
C2 - 8898108
AN - SCOPUS:0030597170
VL - 395
SP - 262
EP - 266
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 2-3
ER -