Recombinant proteins representing full-length and truncated forms of the human immunodeficiency virus type 1 envelope protein gp160 were produced in E. coli and sf9 insect cells. These proteins were denatured and reduced as a function of purification. We adsorbed these proteins onto latex microspheres and used the protein-coated particles as a vehicle to present the antigen in vitro to splenic mononuclear cells from immune mice. Recombinant proteins presented on the latex particles induced antigen-specific proliferative responses that were dependent on the antigen concentration. The proliferative responses were similar to those produced against an identical protein used in soluble form and equivalent protein concentrations. Latex microspheres coated with recombinant proteins could also induce precursor cytotoxic T lymphocytes to mature to functional effector cells in vitro. The use of the latex microspheres to present recombinant proteins as antigens allowed for the use of denatured proteins in our assay that were not soluble in aqueous solutions, such as cell culture media. This system of delivering recombinant proteins in vitro should greatly facilitate the use of recombinant proteins in assays involving live cells.
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http://dx.doi.org/10.1016/0022-1759(91)90266-i | DOI Listing |
Proc Natl Acad Sci U S A
January 2025
Department of Chemistry and Chemical Biology, Baker Laboratory, Cornell University, Ithaca, NY 14853.
Ammonia oxidizing archaea (AOA) are among the most abundant microorganisms on earth and are known to be a major source of nitrous oxide (NO) emissions, although biochemical origins of this NO remain unknown. Enzymological details of AOA nitrogen metabolism are broadly unavailable. We report the recombinant expression, purification, and characterization of a multicopper oxidase, Nmar_1354, from the AOA .
View Article and Find Full Text PDFJ Cell Mol Med
January 2025
Institute of Molecular Medicine, Huaqiao University, Quanzhou, China.
Recombinant adeno-associated virus (rAAV) has emerged as one of the best gene delivery vectors for human gene therapy in vivo. However, the clinical efficacy of rAAV gene therapy is often hindered by the host immune response against its transgene products. Endoplasmic reticulum aminopeptidase 1 (ERAP1) is specialised to process peptides presented by class I molecules of major histocompatibility complex.
View Article and Find Full Text PDFPlant Biotechnol J
January 2025
Institute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Austria.
The production of complex multimeric secretory immunoglobulins (SIgA) in Nicotiana benthamiana leaves is challenging, with significant reductions in complete protein assembly and consequently yield, being the most important difficulties. Expanding the physical dimensions of the ER to mimic professional antibody-secreting cells can help to increase yields and promote protein folding and assembly. Here, we expanded the ER in N.
View Article and Find Full Text PDFMol Biol Rep
January 2025
Department of Structural and Molecular Biology, University College London, London, WC1E 6BT, UK.
Background: Ribosomal protein S6 kinase 1 (p70S6K1) is a member of the AGC family of serine/threonine kinases which plays a role in various cellular processes, including protein synthesis, cell growth, and survival. Dysregulation of p70S6K1, characterized by its overexpression and/or hyperactivation, has been implicated in numerous human pathologies, particularly in several types of cancer. Therefore, generating active, recombinant p70S6K1 is critical for investigating its role in cancer biology and for developing novel diagnostic or therapeutic approaches.
View Article and Find Full Text PDFS D Med
October 2024
Department of Internal Medicine, University of South Dakota Sanford School of Medicine, Sioux Falls, South Dakota.
Acute pericarditis, the predominant pericardial disease, often lacks a clear etiology, with 15-30% of patients experiencing recurrence, rising to 20-50% in those with prior relapses. Autoimmune mechanisms significantly contribute to recurrence, with interleukin-1 identified as a pivotal inflammatory mediator. While NSAIDs, colchicine, and steroids remain staples for acute cases, the spotlight in recurrent pericarditis management has shifted toward immunosuppressive medications.
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