کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1994386 1064775 2008 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Assays for transfer RNA-dependent amino acid biosynthesis
کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Assays for transfer RNA-dependent amino acid biosynthesis
چکیده انگلیسی

Selenocysteinyl-tRNASec, cysteinyl-tRNACys, glutaminyl-tRNAGln, and asparaginyl-tRNAAsn in many organisms are formed in an indirect pathway in which a non-cognate amino acid is first attached to the tRNA. This non-cognate amino acid is then converted to the cognate amino acid by a tRNA-dependent modifying enzyme. The in vitro characterization of these modifying enzymes is challenging due to the fact the substrate, aminoacyl-tRNA, is labile and requires a prior enzymatic step to be synthesized. The need to separate product aa-tRNA from unreacted substrate is typically a labor- and time-intensive task; this adds another impediment in the investigation of these enzymes. Here, we review four different approaches for studying these tRNA-dependent amino acid modifications. In addition, we describe in detail a [32P]/nuclease P1 assay for glutaminyl-tRNAGln and asparaginyl-tRNAAsn formation which is sensitive, enables monitoring of the aminoacyl state of the tRNA, and is less time consuming than some of the other techniques. This [32P]/nuclease P1 method should be adaptable to studying tRNA-dependent selenocysteine and cysteine synthesis.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Methods - Volume 44, Issue 2, February 2008, Pages 139–145
نویسندگان
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