Expansion of the sortase-mediated labeling method for site-specific N-terminal labeling of cell surface proteins on living cells.
نویسندگان
چکیده
The sortase-mediated labeling method, whose applications have been limited to C-terminal labeling of cell surface proteins, has been expanded for site-specific N-terminal labeling of cell surface proteins.
منابع مشابه
Generation of Ca2+-independent sortase A mutants with enhanced activity for protein and cell surface labeling
Sortase A, a calcium-dependent transpeptidase derived from Staphylococcus aureus, is used in a broad range of applications, such as the conjugation of fluorescent dyes and other moieties to proteins or to the surface of eukaryotic cells. In vivo and cell-based applications of sortase have been somewhat limited by the large range of calcium concentrations, as well as by the often transient natur...
متن کاملSite-Specific N- and C-Terminal Labeling of a Single Polypeptide Using Sortases of Different Specificity
The unique reactivity of two sortase enzymes, SrtA(staph) from Staphylococcus aureus and SrtA(strep) from Streptococcus pyogenes, is exploited for site-specific labeling of a single polypeptide with different labels at its N and C termini. SrtA(strep) is used to label the protein's C terminus at an LPXTG site with a fluorescently labeled dialanine nucleophile. Selective N-terminal labeling of p...
متن کاملProtein C-Terminal Labeling and Biotinylation Using Synthetic Peptide and Split-Intein
BACKGROUND Site-specific protein labeling or modification can facilitate the characterization of proteins with respect to their structure, folding, and interaction with other proteins. However, current methods of site-specific protein labeling are few and with limitations, therefore new methods are needed to satisfy the increasing need and sophistications of protein labeling. METHODOLOGY A me...
متن کاملThe first experience of stem cell labeling in Iran using 111In- Oxine [Persian]
Introduction: The field of stem cell biology and regenerative medicine is rapidly moving toward translation to clinical practice. Stem cell therapy seems to be a new treatment option for some diseases. So, tracking the distribution of stem cells is crucial to their therapeutic use. Based on this fact we labeled human mesenchymal stem cells (MSCs) with 111...
متن کاملSite-Specific Chemoenzymatic Labeling of Aerolysin Enables the Identification of New Aerolysin Receptors
Aerolysin is a secreted bacterial toxin that perforates the plasma membrane of a target cell with lethal consequences. Previously explored native and epitope-tagged forms of the toxin do not allow site-specific modification of the mature toxin with a probe of choice. We explore sortase-mediated transpeptidation reactions (sortagging) to install fluorophores and biotin at three distinct sites in...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Chemical communications
دوره 9 شماره
صفحات -
تاریخ انتشار 2009