通过肽阵列分析精氨酰转移酶活性和特异性

Assaying for Arginyltransferase Activity and Specificity by Peptide Arrays.

作者信息

Wang Junling, Kashina Anna S

机构信息

Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.

Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.

出版信息

Methods Mol Biol. 2023;2620:123-127. doi: 10.1007/978-1-0716-2942-0_17.

DOI:10.1007/978-1-0716-2942-0_17

PMID:37010758

Abstract

Here, we describe arginylation assays performed on peptide arrays immobilized on cellulose membranes via chemical synthesis. In this assay, it is possible to simultaneously compare arginylation activity on hundreds of peptide substrates to analyze the specificity of arginyltransferase ATE1 toward its target site(s) and the amino acid sequence context. This assay was successfully employed in prior studies to dissect the arginylation consensus site and enable predictions of arginylated proteins encoded in eukaryotic genomes.

摘要

在此，我们描述了通过化学合成在固定于纤维素膜上的肽阵列上进行的精氨酰化测定。在该测定中，可以同时比较数百种肽底物上的精氨酰化活性，以分析精氨酰转移酶ATE1对其靶位点和氨基酸序列背景的特异性。该测定已在先前的研究中成功用于剖析精氨酰化共有位点，并能够预测真核基因组中编码的精氨酰化蛋白。