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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2018.1
- 수록면
- 322 - 327 (6page)
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초록· 키워드
오류제보하기
Post-translational modification of proteins regulates signaling cascades in eukaryotic system, including plants. Among these modifications, phosphorylation plays an important role in modulating the functional properties of proteins. Plants perceive environmental cues that directly affect the phosphorylation status of many target proteins. To determine the effect of environmentally induced phosphorylation in plants, in vivo methods must be developed. Various in vitro methods are available but, unlike in animals, there is no optimized methodology for detecting protein phosphorylation in planta. Therefore, in this study, a robust, and easy to handle Phos-Tag Mobility Shift Assay (PTMSA) is developed for the in vivo detection of protein phosphorylation in plants by empirical optimization of methods previously developed for animals. Initially, the detection of the phosphorylation status of target proteins using protocols directly adapted from animals failed. Therefore, we optimized the steps in the protocol, from protein migration to the transfer of proteins to PVDF membrane. Supplementing the electrophoresis running buffer with 5 mM NaHSO3 solved most of the problems in protein migration and transfer. The optimization of a fast and robust protocol that efficiently detects the phosphorylation status of plant proteins was successful. This protocol will be a valuable tool for plant scientists interested in the study of protein phosphorylation.
목차
등록된 정보가 없습니다.
참고문헌 (27)
참고문헌 신청
Bruno D / 2006 / Mass Spectrometry and Protein Analysis / Science 312:212~217
James H / 2006 / Assay of protein kinases using radiolabeled ATP: a protocol / Nature Protocols 1:968~971
Chrambach A / 1985 / The practice of quantitative Gel electrophoresis / VCH publishers :177~186
Degeng W / 2003 / Systematic Trans-Genomic Comparison of Protein Kinases between Arabidopsis and Saccharomyces cerevisiae / Plant Physiol 132:2152~2165
Eiji K / 2009 / Separation and detection of large phosphoproteins using Phos-tag SDS-PAGE / Nature Protocols 10:1513~1521
James H / 2006 / Assay of protein kinases using radiolabeled ATP: a protocol / Nature Protocols 1:968~971
Chrambach A / 1985 / The practice of quantitative Gel electrophoresis / VCH publishers :177~186
Degeng W / 2003 / Systematic Trans-Genomic Comparison of Protein Kinases between Arabidopsis and Saccharomyces cerevisiae / Plant Physiol 132:2152~2165
Eiji K / 2009 / Separation and detection of large phosphoproteins using Phos-tag SDS-PAGE / Nature Protocols 10:1513~1521
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