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Chinese Hamster Ovary cell proteins co-purified with recombinant HIV-1 envelopes, part 1

ABSTRACT: Chinese Hamster Ovary (CHO) cells are frequently used to produce recombinant HIV envelope protein gp120. In this study, we characterized the CHO proteins that become co-purified with gp120 when lectin affinity chromatography is used. Many of co-purified CHO proteins identified were extracellular matrix components. We verified a method for separating gp120 from these CHO proteins with anion exchange chromatography, and assessed the biochemical activity of gp120 preparations with and without co-purified CHO proteins.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Cricetulus Griseus (chinese Hamster) (cricetulus Barabensis Griseus)

SUBMITTER: David Plotnik

LAB HEAD: Shiu-Lok Hu

PROVIDER: PXD006060 | Pride | 2017-09-04

REPOSITORIES: Pride

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Extracellular Matrix Proteins Mediate HIV-1 gp120 Interactions with α4β7.

Plotnik David D Guo Wenjin W Cleveland Brad B von Haller Priska P Eng Jimmy K JK Guttman Miklos M Lee Kelly K KK Arthos James J Hu Shiu-Lok SL

Journal of virology 20171013 21

Gut-homing α4β7high CD4+ T lymphocytes have been shown to be preferentially targeted by human immunodeficiency virus type 1 (HIV-1) and are implicated in HIV-1 pathogenesis. Previous studies demonstrated that HIV-1 envelope protein gp120 binds and signals through α4β7 and that this likely contributes to the infection of α4β7high T cells and promotes cell-to-cell virus transmission. Structures withi ...[more]

PMID: 28814519

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Chinese Hamster Ovary cell proteins co-purified with recombinant HIV-1 envelopes, part 2

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Mitochondrial interactome quantitation reveals structural changes in metabolic machinery in failing murine heart

Project description:Advancements of cross-linking mass spectrometry (XL-MS) for structural analysis of proteins bridges the gap between purified systems and native tissue environments. Here, we utilize isobaric quantitative protein interaction reporter technology (iqPIR) to further extend XL-MS to the first system-wide comparative study of mitochondrial proteins from healthy and diseased murine hearts. The failing heart interactome includes 544 statistically significant cross-linked peptide pairs altered in disease condition. Structural insight into ketone oxidation metabolons, OXPHOS machinery, and nucleotide transporter hybrid-conformations, support mitochondrial remodeling in failing heart while bringing forth new hypotheses for pathological mechanisms. Application of quantitative cross-linking technology in live tissue provides molecular-level insight to complex biological systems difficult to model in cell culture, thus providing a valuable resource for study of human diseases.

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AP-MS analysis of human FUBIeS30/FAU WT and non-cleavable G73,4A and G74V mutants

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HIV-1 gp120 impairs B cell proliferation by inducing TGF-β1 production and FcRL4 expression via an α4β7-dependent mechanism

Project description:The anti-HIV humoral immune response following acute infection is delayed and ineffective. HIV envelope protein gp120 binds to and signals through α4β7 on T cells. We show that gp120 also binds and signals through α4β7 on B cells, resulting in an abortive proliferative response. In primary B cells, gp120 signaling through α4β7 resulted in increased expression of TGF-β1 and the B cell inhibitory receptor FcRL4. Co-culture of B cells with HIV-infected autologous CD4+ T cells also resulted in increased B cell FcRL4 expression. These findings indicate that, in addition to inducing chronic immune activation, viral proteins can contribute directly to HIV-associated B cell dysfunction, thus providing a mechanism whereby the virus subverts the early HIV-specific humoral immune response. Forty-two samples in two batches were run with different treatments and different time points.

2013-10-17 | E-GEOD-49019 | biostudies-arrayexpress