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| Content Provider | Journal of Biological Chemistry (JBC) |
|---|---|
| Author | Omi, Rie Kurokawa, Suguru Mihara, Hisaaki Hayashi, Hideyuki Goto, Masaru Miyahara, Ikuko Kurihara, Tatsuo Hirotsu, Ken Esaki, Nobuyoshi |
| Abstract | Selenocysteine lyase (SCL) catalyzes the pyridoxal 5′-phosphate-dependent removal of selenium from l-selenocysteine to yield l-alanine. The enzyme is proposed to function in the recycling of the micronutrient selenium from degraded selenoproteins containing selenocysteine residue as an essential component. The enzyme exhibits strict substrate specificity toward l-selenocysteine and no activity to its cognate l-cysteine. However, it remains unclear how the enzyme distinguishes between selenocysteine and cysteine. Here, we present mechanistic studies of selenocysteine lyase from rat. ESI-MS analysis of wild-type and C375A mutant SCL revealed that the catalytic reaction proceeds via the formation of an enzyme-bound selenopersulfide intermediate on the catalytically essential Cys-375 residue. UV-visible spectrum analysis and the crystal structure of SCL complexed with l-cysteine demonstrated that the enzyme reversibly forms a nonproductive adduct with l-cysteine. Cys-375 on the flexible loop directed l-selenocysteine, but not l-cysteine, to the correct position and orientation in the active site to initiate the catalytic reaction. These findings provide, for the first time, the basis for understanding how trace amounts of a selenium-containing substrate is distinguished from excessive amounts of its cognate sulfur-containing compound in a biological system. |
| Related Links | http://www.jbc.org/content/285/16/12133.abstract |
| Ending Page | 12139 |
| Starting Page | 12133 |
| Page Count | 7 |
| File Format | HTM / HTML PDF |
| ISSN | 00219258 |
| Journal | Journal of Biological Chemistry (JBC) |
| Issue Number | 16 |
| Volume Number | 285 |
| DOI | 10.1074/jbc.M109.084475 |
| e-ISSN | 1083351X |
| Language | English |
| Publisher | American Society for Biochemistry and Molecular Biology |
| Publisher Date | 2010-04-16 |
| Access Restriction | Open |
| Subject Keyword | Crystal Structure Enzyme Mechanisms Enzyme Structure Selenium Sulfur Protein Structure and Folding |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology |
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