Cysteine-free non-canonical C-intein for versatile protein C-terminal labeling through trans -splicing
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| 008 | 210128e20151001xx s 000 0 eng | ||
| 024 | 7 | 0 | |a 10.1007/s00253-015-6796-6 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-015-6796-6 | ||
| 245 | 0 | 0 | |a Cysteine-free non-canonical C-intein for versatile protein C-terminal labeling through trans -splicing |h [Elektronische Daten] |c [Xudong Dai, Qijing Xun, Xiang-Qin Liu, Qing Meng] |
| 520 | 3 | |a Site-specific protein labeling are powerful means of protein research and engineering; however, new and improved labeling methods are greatly needed. Split inteins catalyze a protein trans-splicing reaction that can be used for enzymatic and nearly seamless protein labeling. Non-canonical S11 split intein has been used in an earlier method of protein C-terminal labeling; however, its relatively large (~150 aa) N-intein fused to the target protein often hindered protein expression, folding, and solubility. To solve this problem, here, we have designed and demonstrated a new method of protein C-terminal labeling, by first engineering a functional non-canonical S1 split intein that has an extremely small (12 aa) N-intein and a cysteine-free C-intein. An engineered Rma DnaB S1 split intein was modified to have a cysteine-free C-intein, while still retaining its robust trans-splicing function, which permitted the C-extein in a C-precursor to have a single cysteine for easy and specific linkage with desired labeling groups. The resulting new and generally useful method has two unique advantages: (1) The extremely small (12 aa) N-intein, which must be fused to the C terminus of the target protein, is less likely to hinder the protein expression, folding, and solubility; and (2) the single cysteine in the C-extein may be readily linked to a variety of labeling or modification groups using commercially available reagents. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2015 | ||
| 690 | 7 | |a Protein labeling |2 nationallicence | |
| 690 | 7 | |a Split intein |2 nationallicence | |
| 690 | 7 | |a trans -splicing |2 nationallicence | |
| 690 | 7 | |a Site-specific modifications |2 nationallicence | |
| 700 | 1 | |a Dai |D Xudong |u College of Textiles, Donghua University, 201620, Shanghai, China |4 aut | |
| 700 | 1 | |a Xun |D Qijing |u College of Textiles, Donghua University, 201620, Shanghai, China |4 aut | |
| 700 | 1 | |a Liu |D Xiang-Qin |u Department of Biochemistry & Molecular Biology, Dalhousie University, B3H 4R2, Halifax, Nova Scotia, Canada |4 aut | |
| 700 | 1 | |a Meng |D Qing |u College of Textiles, Donghua University, 201620, Shanghai, China |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/19(2015-10-01), 8151-8161 |x 0175-7598 |q 99:19<8151 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-015-6796-6 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-015-6796-6 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Dai |D Xudong |u College of Textiles, Donghua University, 201620, Shanghai, China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Xun |D Qijing |u College of Textiles, Donghua University, 201620, Shanghai, China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Liu |D Xiang-Qin |u Department of Biochemistry & Molecular Biology, Dalhousie University, B3H 4R2, Halifax, Nova Scotia, Canada |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Meng |D Qing |u College of Textiles, Donghua University, 201620, Shanghai, China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/19(2015-10-01), 8151-8161 |x 0175-7598 |q 99:19<8151 |1 2015 |2 99 |o 253 | ||