MassIVE MSV000079886

Partial Public PXD004519

MudPIT analysis of HSV-1 infected and mock-infected microsomal membranes and nuclear envelopes

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Description

Nuclear envelopes (NEs) and microsomal membranes (MMs) were isolated from HeLa cells infected or not with HSV-1 using well-established procedures as described in: Wilkie GS, et al. (2011) Mol Cell Proteomics 10: M110 003129. Korfali N, et al. (2010) Mol Cell Proteomics 9: 2571-2585. Florens L, Korfali N, Schirmer EC (2008) Methods Mol Biol 432: 117-137. Membrane pellets were denatured, reduced, alkylated then digested with endoproteinase LysC followed by trypsin. Peptides were analyzed by Multidimensional Protein Identification Technology (MudPIT) as described in: Florens L, Washburn MP (2006) Methods Mol Biol 328: 159-175. RAW files were extracted into ms2 file format using RawDistiller v. 1.0 as in: Zhang, Y.; Wen, Z.; Washburn, M. P.; Florens, L. (2011) Anal Chem, 83, (24), 9344-51. MS/MS spectra were queried for peptide sequence information using SEQUEST v.27 (rev.9) as in: Eng J, McCormack A, Yates Jr (1994) J Am Soc Mass Spectrom 5: 976-989. Results from different runs were compared using DTASelect and CONTRAST as in: Tabb DL, McDonald WH, Yates JR (2002) J Proteome Res 1: 21-26. To estimate relative protein levels, distributed normalized spectral abundance factors (dNSAFs) were calculated for each non-redundant protein as in: Zhang Y, Wen Z, Washburn MP, Florens L (2010) Anal Chem 82: 2272-2281. [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: nuclear envelope ; herpesvirus ; nuclear egress ; vesicle fusion protein

Contact

Principal Investigators:
(in alphabetical order) 		Laurence Florens
Submitting User: laflorens

Publications

Saiz-Ros N, Czapiewski R, Epifano I, Stevenson A, Swanson SK, Dixon CR, Zamora DB, McElwee M, Vijayakrishnan S, Richardson CA, Dong L, Kelly DA, Pytowski L, Goldberg MW, Florens L, Graham SV, Schirmer EC.
Host Vesicle Fusion Protein VAPB Contributes to the Nuclear Egress Stage of Herpes Simplex Virus Type-1 (HSV-1) Replication.
Cells. Epub 2019 Feb 3.

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