ArticleThe Journal of biological chemistry2009
Purification and reconstitution of the antigen transport complex TAP: a prerequisite for determination of peptide stoichiometry and ATP hydrolysis.
Article in The Journal of biological chemistry, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed.
- A Structural Bioinformatics-Guided Study of Adenosine Triphosphate-Binding Cassette (ABC) Transporters and Their Substrates.Membranes · 2025Article
- Semisynthetic Viral Inhibitor for Light Control of the MHC I Peptide Loading Complex.Angewandte Chemie (International ed. in English) · 2022Article
- Determining small-molecule permeation through lipid membranes.Nature protocols · 2022Review
- The lysosomal transporter TAPL has a dual role as peptide translocator and phosphatidylserine floppase.Nature communications · 2022Article
- Functional Overexpression of Membrane Proteins in E. coli: The Good, the Bad, and the Ugly.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Quantitative real-time analysis of the efflux by the MacAB-TolC tripartite efflux pump clarifies the role of ATP hydrolysis within mechanotransmission mechanism.Communications biology · 2021Article
- Substrate recognition and ATPase activity of theThe Journal of biological chemistry · 2020Article
- Peptide translocation by the lysosomal ABC transporter TAPL is regulated by coupling efficiency and activation energy.Scientific reports · 2019Article
- Towards Identification of the Substrates of ATP-Binding Cassette Transporters.Plant physiology · 2018Review
- Moving the Cellular Peptidome by Transporters.Frontiers in cell and developmental biology · 2018Review
- Activity of the purified plant ABC transporter NtPDR1 is stimulated by diterpenes and sesquiterpenes involved in constitutive and induced defenses.The Journal of biological chemistry · 2017Article
- Structure and Dynamics of Antigenic Peptides in Complex with TAP.Frontiers in immunology · 2017Review
- Ultrasensitive quantification of TAP-dependent antigen compartmentalization in scarce primary immune cell subsets.Nature communications · 2015Article
- Unexpected lack of specificity of a rabbit polyclonal TAP-L (ABCB9) antibody.F1000Research · 2015Article
- A negative feedback modulator of antigen processing evolved from a frameshift in the cowpox virus genome.PLoS pathogens · 2014Article
- An annular lipid belt is essential for allosteric coupling and viral inhibition of the antigen translocation complex TAP (transporter associated with antigen processing).The Journal of biological chemistry · 2014Article
- Mechanistic determinants of the directionality and energetics of active export by a heterodimeric ABC transporter.Nature communications · 2014Article
- Structural basis for heavy metal detoxification by an Atm1-type ABC exporter.Science (New York, N.Y.) · 2014Article
- Differential phospholipid substrates and directional transport by ATP-binding cassette proteins ABCA1, ABCA7, and ABCA4 and disease-causing mutants.The Journal of biological chemistry · 2013Article
- Structures of ABCB10, a human ATP-binding cassette transporter in apo- and nucleotide-bound states.Proceedings of the National Academy of Sciences of the United States of America · 2013Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The transporter associated with antigen processing (TAP) is an essential machine of the adaptive immune system that translocates antigenic peptides from the cytosol into the endoplasmic reticulum lumen for loading of major histocompatibility class I molecules. To examine this ABC transport complex in mechanistic detail, we have established, after extensive screening and optimization, the solubilization, purification, and reconstitution for TAP to preserve its function in each step. This allowed us to determine the substrate-binding stoichiometry of the TAP complex by fluorescence cross-correlation spectroscopy. In addition, the TAP complex shows strict coupling between peptide binding and ATP hydrolysis, revealing no basal ATPase activity in the absence of peptides. These results represent an optimal starting point for detailed mechanistic studies of the transport cycle of TAP by single molecule experiments to analyze single steps of peptide translocation and the stoichiometry between peptide transport and ATP hydrolysis.
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