Biology:Tobacco acid pyrophosphatase
Tobacco Acid Pyrophosphatase (TAP) is an enzyme that catalyses the hydrolysis of a phosphoric ester bond in a broad spectrum of molecules,[1] including the 5'-end of mRNA.
During mRNA maturation the 5' triphosphate of the new mRNA molecule is rapidly removed. The diphosphate 5' end then attacks the α-phosphorus atom of a methylated GTP to form a very unusual 5'-5' triphosphate linkage, called cap.[2] In molecular biology, TAP is used to hydrolyse a phosphodiester bond of this particular structure and release a mRNA molecule with only one phosphate group in the 5'-end,[3] for instance in protocols for RACE (Rapid Amplification of cDNA Ends).
Commercial production of TAP was discontinued in 2015, however Cap-Clip acid pyrophosphatase has been found to perform nearly identically in genome-wide studies.[4]
References
- ↑ "A novel phosphodiesterase from cultured tobacco cells". Biochemistry 15 (10): 2185–90. 1976. doi:10.1021/bi00655a024. PMID 6041.
- ↑ JEREMY M. BERG; JOHN L. TYMOCZKOL; LUBERT STRYER (2002). Biochemistry (28.3.1 ed.). https://archive.org/details/biochemistry200100jere.Online version
- ↑ Lockard, RE; Rieser, L; Vournakis JN. (1981). "Labeling of eukaryotic messenger RNA 5' terminus with phosphorus -32: use of tobacco acid pyrophosphatase for removal of cap structures". Gene Amplif Anal 2: 229–51. PMID 6152889.
- ↑ Pelechano, Vicent; Wei, Wu; Steinmetz, Lars M. (February 2016). "Genome-wide quantification of 5′-phosphorylated mRNA degradation intermediates for analysis of ribosome dynamics" (in en). Nature Protocols 11 (2): 359–376. doi:10.1038/nprot.2016.026. ISSN 1750-2799. PMID 26820793.
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