Dataset Information

Human Adenovirus 5 (HAdV5), LC-MSMS

ABSTRACT: Using high-resolution mass spectrometry (MS)-based proteomics in combination with multiple protease digestion, we profiled, with on average 90% sequence coverage, all 13 viral proteins present in an human adenovirus (HAdV) vector. This in-depth profile provided multiple peptide-based evidence on intrinsic protease activity affecting several HAdV proteins. Next, the generated peptide library was used to develop a targeted proteomics method using selected reaction monitoring (SRM) aimed at quantitative profiling of the stoichiometry of all 13 proteins present in the HAdV. We also used this method to probe the release of specific virus proteins initiated by thermal stimulation, mimicking the early stage of HAdV disassembly during entry into host cells. We confirmed the copy numbers of the most well characterized viral capsid components and established the copy numbers for proteins whose stoichiometry has so far not been accurately defined. We also found that heating HAdV induces the complete release of the penton base and fiber proteins as well as a substantial release of protein VIII and VI. For these latter proteins, maturational proteolysis by the adenoviral protease (AVP) leads to the differential release of fragments with certain peptides being fully released and others largely retained in the AdV particles. This information is likely to be beneficial for the ongoing interpretation of high resolution cryoEM and X-ray electron density maps.

INSTRUMENT(S): LTQ Orbitrap, Q Exactive

ORGANISM(S): Human Adenovirus C Serotype 5 (hadv-5) (human Adenovirus 5)

SUBMITTER: Marco Benevento

LAB HEAD: Albert J.R. Heck

PROVIDER: PXD000591 | Pride | 2014-06-09

REPOSITORIES: pride

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Publications

Adenovirus composition, proteolysis, and disassembly studied by in-depth qualitative and quantitative proteomics.

Benevento Marco M Di Palma Serena S Snijder Joost J Moyer Crystal L CL Reddy Vijay S VS Nemerow Glen R GR Heck Albert J R AJR

The Journal of biological chemistry 20140303 16

Using high-resolution MS-based proteomics in combination with multiple protease digestion, we profiled, with on average 90% sequence coverage, all 13 viral proteins present in an human adenovirus (HAdV) vector. This in-depth profile provided multiple peptide-based evidence on intrinsic protease activity affecting several HAdV proteins. Next, the generated peptide library was used to develop a targeted proteomics method using selected reaction monitoring (SRM) aimed at quantitative profiling of t ...[more]

PMID: 24591515

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	HAdV_1h_Qexactive.raw	Raw
	HAdV_1h_Qexactive_2.raw	Raw
	HAdV_1h_Qexactive_3.raw	Raw
	HAdV_90m_Qexactive.raw	Raw
	HAdV_90m_Qexactive_2.raw	Raw