Isolation and characterization of a nitric oxide synthase (NOS)-like protein of pea (Pisum sativum L.)
Nitric oxide synthase (NOS) activity based on citrulline formation assay, which was used in mammalian system, was detected in Pisum Sativum L. (pea) extracts. The pea NOS-like protein was most efficiently extracted with the addition of protease inhibitors (ethylene bis (oxyethylenenitrilo)tetraaceti...
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| Format: | Article |
| Language: | English |
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Penerbit Universiti Sains Malaysia
2007
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| Online Access: | http://psasir.upm.edu.my/id/eprint/51944/ http://psasir.upm.edu.my/id/eprint/51944/1/Isolation%20and%20characterization%20of%20a%20nitric%20oxide%20synthase%20%28NOS%29-like%20protein%20of%20pea%20%28Pisum%20sativum%20L.%29.pdf |
| _version_ | 1848851971206807552 |
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| author | Wong, Mui Yun Huang, Jeng Sheng Davis, Eric L. |
| author_facet | Wong, Mui Yun Huang, Jeng Sheng Davis, Eric L. |
| author_sort | Wong, Mui Yun |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Nitric oxide synthase (NOS) activity based on citrulline formation assay, which was used in mammalian system, was detected in Pisum Sativum L. (pea) extracts. The pea NOS-like protein was most efficiently extracted with the addition of protease inhibitors (ethylene bis (oxyethylenenitrilo)tetraacetic acid (EGTA) and leupeptin) in the extraction buffer and under alkaline condition (pH 8.5–9.0) as compared to neutral condition in mammalian system. The precipitation of this protein with various concentrations of ammonium sulfate, sodium citrate and sodium chloride caused rapid loss of NOS activity, in contrast to that in the mammalian system, and the protein was not precipitated by organic solvents (acetone or polyethylene glycol, PEG). The pea NOS-like protein was successfully isolated using ion-exchange column, but did not bind to β-nicotinamide adenine dinucleotide phosphate (NADPH) and calmodulin affinity columns suggesting that it lacked binding sites for the cofactors NADPH and calmodulin that were required for NOS activity in mammalian cells. The results indicated that the pea NOS like protein was significantly different in structure from mammalian NOS. |
| first_indexed | 2025-11-15T10:30:39Z |
| format | Article |
| id | upm-51944 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T10:30:39Z |
| publishDate | 2007 |
| publisher | Penerbit Universiti Sains Malaysia |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-519442017-05-04T04:28:23Z http://psasir.upm.edu.my/id/eprint/51944/ Isolation and characterization of a nitric oxide synthase (NOS)-like protein of pea (Pisum sativum L.) Wong, Mui Yun Huang, Jeng Sheng Davis, Eric L. Nitric oxide synthase (NOS) activity based on citrulline formation assay, which was used in mammalian system, was detected in Pisum Sativum L. (pea) extracts. The pea NOS-like protein was most efficiently extracted with the addition of protease inhibitors (ethylene bis (oxyethylenenitrilo)tetraacetic acid (EGTA) and leupeptin) in the extraction buffer and under alkaline condition (pH 8.5–9.0) as compared to neutral condition in mammalian system. The precipitation of this protein with various concentrations of ammonium sulfate, sodium citrate and sodium chloride caused rapid loss of NOS activity, in contrast to that in the mammalian system, and the protein was not precipitated by organic solvents (acetone or polyethylene glycol, PEG). The pea NOS-like protein was successfully isolated using ion-exchange column, but did not bind to β-nicotinamide adenine dinucleotide phosphate (NADPH) and calmodulin affinity columns suggesting that it lacked binding sites for the cofactors NADPH and calmodulin that were required for NOS activity in mammalian cells. The results indicated that the pea NOS like protein was significantly different in structure from mammalian NOS. Penerbit Universiti Sains Malaysia 2007 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/51944/1/Isolation%20and%20characterization%20of%20a%20nitric%20oxide%20synthase%20%28NOS%29-like%20protein%20of%20pea%20%28Pisum%20sativum%20L.%29.pdf Wong, Mui Yun and Huang, Jeng Sheng and Davis, Eric L. (2007) Isolation and characterization of a nitric oxide synthase (NOS)-like protein of pea (Pisum sativum L.). Journal of Bioscience, 18 (2). pp. 9-23. ISSN 1985-3718; ESSN: 2180-4249 http://www.tlsr.usm.my/TLSRvol18no2.html |
| spellingShingle | Wong, Mui Yun Huang, Jeng Sheng Davis, Eric L. Isolation and characterization of a nitric oxide synthase (NOS)-like protein of pea (Pisum sativum L.) |
| title | Isolation and characterization of a nitric oxide synthase (NOS)-like protein of pea (Pisum sativum L.) |
| title_full | Isolation and characterization of a nitric oxide synthase (NOS)-like protein of pea (Pisum sativum L.) |
| title_fullStr | Isolation and characterization of a nitric oxide synthase (NOS)-like protein of pea (Pisum sativum L.) |
| title_full_unstemmed | Isolation and characterization of a nitric oxide synthase (NOS)-like protein of pea (Pisum sativum L.) |
| title_short | Isolation and characterization of a nitric oxide synthase (NOS)-like protein of pea (Pisum sativum L.) |
| title_sort | isolation and characterization of a nitric oxide synthase (nos)-like protein of pea (pisum sativum l.) |
| url | http://psasir.upm.edu.my/id/eprint/51944/ http://psasir.upm.edu.my/id/eprint/51944/ http://psasir.upm.edu.my/id/eprint/51944/1/Isolation%20and%20characterization%20of%20a%20nitric%20oxide%20synthase%20%28NOS%29-like%20protein%20of%20pea%20%28Pisum%20sativum%20L.%29.pdf |