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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Shi, Xinghua Jung, Yonil Lin, Li-Jung Liu, Cheng Wu, Cong Cann, Isaac K. O. Ha, Taekjip |
| Description | Country affiliation: United States Author Affiliation: Shi X ( Howard Hughes Medical Institute, Urbana, Illinois, USA.) |
| Abstract | A major hurdle for molecular mechanistic studies of many proteins is the lack of a general method for fluorescence labeling with high efficiency, specificity and speed. By incorporating an aldehyde motif genetically into a protein and improving the labeling kinetics substantially under mild conditions, we achieved fast, site-specific labeling of a protein with â¼100% efficiency while maintaining the biological function. We show that an aldehyde-tagged protein can be specifically labeled in cell extracts without protein purification and then can be used in single-molecule pull-down analysis. We also show the unique power of our method in single-molecule studies on the transient interactions and switching between two quantitatively labeled DNA polymerases on their processivity factor. |
| File Format | HTM / HTML |
| ISSN | 15487091 |
| e-ISSN | 15487105 |
| DOI | 10.1038/nmeth.1954 |
| Journal | Nature Methods |
| Issue Number | 5 |
| Volume Number | 9 |
| Language | English |
| Publisher | Nature Publishing Group |
| Publisher Date | 2012-04-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Discipline Clinical Laboratory Techniques Aldehydes Chemistry Carbocyanines Chemical Synthesis Dna-directed Dna Polymerase Fluorescent Dyes Kinetics Microscopy, Fluorescence Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't Research Support, U.s. Gov't, Non-p.h.s. |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology Biotechnology |
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