Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Pivot Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Target Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Query: EC:4.1.1.6 (
CAD
)
4,420
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Using a newly developed dual-cell quadrupole linear ion trap-orbitrap hybrid mass spectrometer (dcQLT-orbitrap), we demonstrate the utility of collecting high-resolution tandem mass spectral data for large-scale shotgun proteomics. Multiple nanoLC-MS/MS experiments on both an older generation quadrupole linear ion trap-orbitrap hybrid (QLT-orbitrap) and the dcQLT-orbitrap, using both resonant-excitation
CAD
and beam-type
CAD
(
HCD
), were performed. Resulting from various technological advances (e.g., a stacked ring ion guide AP inlet, a dual cell QLT), the dcQLT-orbitrap exhibited increased duty cycle (approximately 1.5-2 times) and sensitivity for both
CAD
(ion trap detection) and
HCD
(orbitrap detection) methods. As compared to the older system, the dcQLT-orbitrap produced significantly more unique peptide identifications for both methods (approximately 30% improvement for
CAD
and approximately 115% improvement for
HCD
). The sizable improvement of the
HCD
method on the dcQLT-orbitrap system outperforms the current standard method of
CAD
with ion trap detection for large-scale analysis. Finally, we demonstrate that the increased
HCD
performance translates to a direct and substantial improvement in protein quantitation precision using isobaric tags.
...
PMID:Analysis of tandem mass spectra by FTMS for improved large-scale proteomics with superior protein quantification. 1993 23
Beam-type collisional activation dissociation (
HCD
) offers many advantages over resonant excitation collision-activated dissociation, including improved identification of phosphorylated peptides and compatibility with isobaric tag-based quantitation (e.g. tandem mass tag (TMT) and iTRAQ). However,
HCD
typically requires specially designed and dedicated collision cells. Here we demonstrate that
HCD
can be performed in the ion injection pathway of a mass spectrometer with a standard atmospheric inlet (iHCD). Testing this method on complex peptide mixtures revealed similar identification rates to collision-activated dissociation (2883 versus 2730 IDs for iHCD/
CAD
, respectively) and precursor-product-conversion efficiency comparable to that achieved within a dedicated collision cell. Compared with pulsed-q dissociation, a quadrupole ion trap-based method that retains low-mass isobaric tag reporter ions, iHCD yielded isobaric tag for relative and absolute quantification reporter ions 10-fold more intense. This method involves no additional hardware and can theoretically be implemented on any mass spectrometer with an atmospheric inlet.
...
PMID:Higher-energy collision-activated dissociation without a dedicated collision cell. 2139 38
Analysis of 15,897 low-energy (
CAD
) and 10,878 higher-energy (
HCD
) collisional dissociation mass spectra of doubly protonated tryptic peptides taken with high resolution revealed that the rate of sequence scrambling due to b-ion cyclization is negligible (<1%) and can be safely ignored as a possible source of erroneous sequence assignment in shotgun proteomics. On the other hand, there is significant presence of normal (non-scrambled) internal fragments in
HCD
, which should be taken into account by MS/MS search engines.
...
PMID:Sequence scrambling in shotgun proteomics is negligible. 2195 93