Recombinant expression, characterization, and quantification in human cancer cell lines of the Anaplastic Large-Cell Lymphoma-characteristic NPM-ALK fusion protein.
Katerina Kourentzi,Mary Crum,Ujwal Patil,Ana Prebisch,Dimple Chavan,Binh Vu,Zihua Zeng,Dmitri Litvinov,Youli Zu,Richard C. Willson,Richard C. Willson +10 more
TL;DR: It is estimated that N PM-ALK fusion protein is expressed at substantial levels in both Karpas 299 and SU-DHL-1 cells and compared NPM-ALK/ β-actin ratios determined by ELISA to those independently determined by two-dimensional electrophoresis and showed that the two methods are in good agreement.
read more
Abstract: Systemic anaplastic large cell lymphoma (ALCL) is an aggressive T-cell lymphoma most commonly seen in children and young adults. The majority of pediatric ALCLs are associated with the t(2;5)(p23;q35) translocation which fuses the Anaplastic Lymphoma Kinase (ALK) gene with the Nucleophosmin (NPM) gene. The NPM-ALK fusion protein is a constitutively-active tyrosine kinase, and plays a major role in tumor pathogenesis. In an effort to advance novel diagnostic approaches and the understanding of the function of this fusion protein in cancer cells, we expressed in E. coli, purified and characterized human NPM-ALK fusion protein to be used as a standard for estimating expression levels in cultured human ALCL cells, a key tool in ALCL pathobiology research. We estimated that NPM-ALK fusion protein is expressed at substantial levels in both Karpas 299 and SU-DHL-1 cells (ca. 4–6 million molecules or 0.5–0.7 pg protein per cell; based on our in-house developed NPM-ALK ELISA; LOD of 40 pM) as compared to the ubiquitous β-actin protein (ca. 64 million molecules or 4.5 pg per lymphocyte). We also compared NPM-ALK/ β-actin ratios determined by ELISA to those independently determined by two-dimensional electrophoresis and showed that the two methods are in good agreement.
read more
Chat with Paper
AI Agents for this Paper
Find similar papers on Google Scholar, PubMed and Arxiv
Write a critical review of this paper
Analyze citations of this paper to find unaddressed research gaps
Citations
Effect of Diet and Oxidative Stress in the Pathogenesis of Lymphoproliferative Disorders
TL;DR: In vitro and in vivo studies on the possible interaction between lymphomas, oxidative stress, and diet are investigated and a variety of dietary regimens and substances introduced with the diet that may have antioxidant and antiproliferative effects were assessed.
6
Selective Elimination of Senescent Cancer Cells by Galacto-Modified PROTACs.
Feng Gao,Giri R. Gnawali,Hang Xu,Yue Dong,Xiang Meng,Wenpan Li,Zhiren Wang,Byrdie Lopez,Jennifer S. Carew,Steffan T. Nawrocki,Jianqin Lu,Qing-Yu Zhang,Wei Wang +12 more
TL;DR: A new strategy for selective degradation of target proteins in senescent cancer cells is devised that utilizes a prodrug composed of the SA-β-gal substrate galactose (galacto) and the proteolysis-targeting chimeras (PROTACs) as senolytic agents.
6
Comparative analysis of protein expression systems and PTM landscape in the study of transcription factor ELK-1.
TL;DR: In this article , the ETS transcription factor ELK-1 serves as a paradigm for cellular responses to mitogens and can synergise with androgen receptor to promote prostate cancer progression, although in vitro protein function analyses to date have largely overlooked its complex PTM landscapes.
3
The involvement of oxidative stress in non-Hodgkin’s lymphomas; a review of the literature
10 Apr 2022
TL;DR: In this article , a review summarizes the latest findings in understanding the ROS-linked signaling pathways in the initiation of lymphomagenesis, disease progression, metastasis, as well as in the pharmacodynamics of specific treatments for this malignancy.
1
References
Actin and tubulin of normal and leukaemic lymphocytes.
H Atkins,P J Anderson +1 more
TL;DR: Double labelling and the isolation of actin- and tubulin-derived peptides were used to determine the amounts of these proteins in peripheral lymphocytes from normal donors and from patients with chronic lymphocytic leukaemia.
Lysis of Cultured Cells for Immunoprecipitation
TL;DR: This protocol presents separate methods for lysing cells grown as monolayer cultures and for cells grown in suspension, which can be gentler to the protein of interest than mechanical homogenization methods.