Unknown

Dataset Information

0

Top-down protein identification of proteasome proteins with nanoLC-FT-ICR-MS employing data-independent fragmentation methods.


ABSTRACT: A comparison of different data-independent fragmentation methods combined with LC coupled to high-resolution FT-ICR-MS/MS is presented for top-down MS of protein mixtures. Proteins composing the 20S and 19S proteasome complexes and their PTMs were identified using a 15 T FT-ICR mass spectrometer. The data-independent fragmentation modes with LC timescales allowed for higher duty-cycle measurements that better suit online LC-FT-ICR-MS. Protein top-down dissociation was effected by funnel-skimmer collisionally activated dissociation (FS-CAD) and CASI (continuous accumulation of selected ions)-CAD. The N-termini for 9 of the 14 20S proteasome proteins were found to be modified, and the ?3 protein was found to be phosphorylated; these results are consistent with previous reports. Mass-measurement accuracy with the LC-FT-ICR system for the 20- to 30-kDa 20S proteasome proteins was 1 ppm. The intact mass of the 100-kDa Rpn1 subunit from the 19S proteasome complex regulatory particle was measured with a deviation of 17 ppm. The CASI-CAD technique is a complementary tool for intact-protein fragmentation and is an effective addition to the growing inventory of dissociation methods that are compatible with online protein separation coupled to FT-ICR-MS.

SUBMITTER: Lakshmanan R 

PROVIDER: S-EPMC4045009 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature

altmetric image

Publications

Top-down protein identification of proteasome proteins with nanoLC-FT-ICR-MS employing data-independent fragmentation methods.

Lakshmanan Rajeswari R   Wolff Jeremy J JJ   Alvarado Rudy R   Loo Joseph A JA  

Proteomics 20140326 10


A comparison of different data-independent fragmentation methods combined with LC coupled to high-resolution FT-ICR-MS/MS is presented for top-down MS of protein mixtures. Proteins composing the 20S and 19S proteasome complexes and their PTMs were identified using a 15 T FT-ICR mass spectrometer. The data-independent fragmentation modes with LC timescales allowed for higher duty-cycle measurements that better suit online LC-FT-ICR-MS. Protein top-down dissociation was effected by funnel-skimmer  ...[more]

Similar Datasets

| S-EPMC5353847 | biostudies-literature
| S-EPMC4095805 | biostudies-literature
| S-EPMC10557258 | biostudies-literature
| S-EPMC3479637 | biostudies-literature
| S-EPMC8059610 | biostudies-literature
| S-EPMC4144182 | biostudies-literature
| S-EPMC3712280 | biostudies-literature
| S-EPMC3411821 | biostudies-literature
| S-EPMC5827162 | biostudies-literature
| S-EPMC5661887 | biostudies-literature