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

ABSTRACT: 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.

INSTRUMENT(S): LTQ

ORGANISM(S): Homo Sapiens (ncbitaxon:9606) Herpes Simplex Virus (type 1 / Strain 17) (ncbitaxon:10299)

SUBMITTER: Laurence Florens 

PROVIDER: MSV000079886 | MassIVE | Tue Jul 05 07:55:00 BST 2016

SECONDARY ACCESSION(S): PXD004519

REPOSITORIES: MassIVE