Identification of ARTS-1 as a novel TNFR1-binding protein that promotes TNFR1 ectodomain shedding
Xinle Cui, Feras Hawari, Sura Alsaaty, Marion Lawrence, Christian A. Combs, Weidong Geng, Farshid N. Rouhani, Dianne Miskinis, Stewart J. Levine
Xinle Cui, Feras Hawari, Sura Alsaaty, Marion Lawrence, Christian A. Combs, Weidong Geng, Farshid N. Rouhani, Dianne Miskinis, Stewart J. Levine
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Article

Identification of ARTS-1 as a novel TNFR1-binding protein that promotes TNFR1 ectodomain shedding

Abstract

Research Article

Authors

Xinle Cui, Feras Hawari, Sura Alsaaty, Marion Lawrence, Christian A. Combs, Weidong Geng, Farshid N. Rouhani, Dianne Miskinis, Stewart J. Levine

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Characterization of GST–ARTS-1 aminopeptidase activity. (a) Generation o...
Characterization of GST–ARTS-1 aminopeptidase activity. (a) Generation of GST–ARTS-1. Soluble and insoluble protein fractions were isolated from BL21 E. coli transformed with empty pGEX-6P-1 (Lane 1, soluble fraction; Lane 2, insoluble fraction) or ARTS-1 pGEX-6P-1 (Lane 3, soluble fraction; Lane 4, insoluble fraction). Proteins were subjected to SDS-PAGE and stained with Coomassie brilliant blue. Purified GST–ARTS-1 fusion protein from the insoluble fraction is shown as a predominant 130-kDa band in Lane 5, and the 26-kDa purified control GST tag is shown in Lane 6. (b) FPLC analysis of purified recombinant GST–ARTS-1 fusion protein revealed a major peak that eluted at approximately 40 minutes. (c) Assay of GST–ARTS-1 aminopeptidase activity. FPLC fractions were assessed for aminopeptidase activity using a phenylalanine–p-nitroanilide (Phe-pNA) substrate. Phenylalanine aminopeptidase activity was present in pooled fractions eluting from 38 to 44 minutes, which correlated with the major FPLC peak.