Identification of 12-Lipoxygenase Interaction with Cellular Proteins by Yeast Two-Hybrid Screening
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- 作者：Keqin Tang ; Russell L. Finley ; Jr. ; Daotai Nie ; and Kenneth V. Honn
- 刊名：Biochemistry
- 出版年：2000
- 出版时间：March 28, 2000
- 年：2000
- 卷：39
- 期：12
- 页码：3185 - 3191
- 全文大小：356K
- 年卷期：v.39,no.12(March 28, 2000)
- ISSN：1520-4995
文摘
The platelet isoform of 12-lipoxygenase (12-LOX) is expressed in a variety of human tumors.12-LOX metabolizes arachidonic acid to 12(S)-hydroxyeicosateraenoic acid (12(S)-HETE), which inducesa number of cellular responses associated with tumor progression and metastasis. Little is known about12-LOX regulation and no direct regulators of 12-LOX activity have been identified. To identify potentialregulators of 12-LOX, we isolated cDNAs encoding 12-LOX interacting proteins using the yeast two-hybrid system. We screened a yeast two-hybrid interaction library from human epidermoid carcinomaA431 cells and identified four cellular proteins that interact specifically with 12-LOX. We identified typeII keratin 5, lamin A, the cytoplasmic domain of integrin 
4 subunit and a phosphoprotein C8FW as12-LOX interacting proteins. Here, we demonstrated that keratin 5, a 58 kD protein required for formationof 8 nm intermediate filaments, binds to 12-LOX in human tumor cells and may contribute to the regulatedtrafficking of 12-LOX. We also showed that lamin A binds 12-LOX in human tumor cells. These proteinsprovide the first candidate regulators of 12-LOX.
4 subunit and a phosphoprotein C8FW as12-LOX interacting proteins. Here, we demonstrated that keratin 5, a 58 kD protein required for formationof 8 nm intermediate filaments, binds to 12-LOX in human tumor cells and may contribute to the regulatedtrafficking of 12-LOX. We also showed that lamin A binds 12-LOX in human tumor cells. These proteinsprovide the first candidate regulators of 12-LOX.