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Applications of a matrix-assisted laser desorption/ionization orthogonal time-of-flight mass spectrometer. l. Metastable decay and collision-induced dissociation for sequencing peptides.
Rapid Commun Mass Spectrom. 2005; 19(2):213-20.RC

Abstract

The use of a high-performance orthogonal time-of-flight (o-TOF) mass spectrometer for sequence analysis is described. The mass spectrometer is equipped with a matrix-assisted laser desorption/ionization (MALDI) source that operates at elevated pressure, 0.01-1 Torr. Ion fragmentation is controlled by varying the pressure of the buffer gas, the laser energy, the voltage difference between the MALDI target and the adjacent sampling cone, and between the cone and the quadrupole ion guide. The peptides were analyzed under optimal ionization conditions to obtain their molecular mass, and under conditions that promote ion dissociation via metastable decomposition or collision-induced dissociation (CID). The fragmentation spectra were used to obtain sequence information. Ion dissociation was promoted via three configurations of the ionization parameters. All methods yielded sequencing-grade b- and y-type ions. Two binary mixtures of peptides were used to demonstrate that: (1) external calibration provides a standard deviation (sigma) of 4 ppm with a mode of 9 ppm; and (2) that peptides with molecular masses that differ by a factor of two may be independently fragmented by appropriately choosing the CID energy and the low-mass cut-off. Analyses of tryptic digests employed liquid chromatography (LC), deposition of the eluant on a target, and finally MALDI-TOF mass spectrometry. The mass fingerprint and the (partial) sequence of the tryptic peptides were matched to their precursor protein via database searches.

Authors+Show Affiliations

MDS Sciex, 71 Four Valley Drive, Concord, Ontario L4K 4V8, Canada. suzanne.ackloo@sciex.comNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15593255

Citation

Ackloo, Suzanne, and Alexandre Loboda. "Applications of a Matrix-assisted Laser Desorption/ionization Orthogonal Time-of-flight Mass Spectrometer. L. Metastable Decay and Collision-induced Dissociation for Sequencing Peptides." Rapid Communications in Mass Spectrometry : RCM, vol. 19, no. 2, 2005, pp. 213-20.
Ackloo S, Loboda A. Applications of a matrix-assisted laser desorption/ionization orthogonal time-of-flight mass spectrometer. l. Metastable decay and collision-induced dissociation for sequencing peptides. Rapid Commun Mass Spectrom. 2005;19(2):213-20.
Ackloo, S., & Loboda, A. (2005). Applications of a matrix-assisted laser desorption/ionization orthogonal time-of-flight mass spectrometer. l. Metastable decay and collision-induced dissociation for sequencing peptides. Rapid Communications in Mass Spectrometry : RCM, 19(2), 213-20.
Ackloo S, Loboda A. Applications of a Matrix-assisted Laser Desorption/ionization Orthogonal Time-of-flight Mass Spectrometer. L. Metastable Decay and Collision-induced Dissociation for Sequencing Peptides. Rapid Commun Mass Spectrom. 2005;19(2):213-20. PubMed PMID: 15593255.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Applications of a matrix-assisted laser desorption/ionization orthogonal time-of-flight mass spectrometer. l. Metastable decay and collision-induced dissociation for sequencing peptides. AU - Ackloo,Suzanne, AU - Loboda,Alexandre, PY - 2004/12/14/pubmed PY - 2005/3/9/medline PY - 2004/12/14/entrez SP - 213 EP - 20 JF - Rapid communications in mass spectrometry : RCM JO - Rapid Commun Mass Spectrom VL - 19 IS - 2 N2 - The use of a high-performance orthogonal time-of-flight (o-TOF) mass spectrometer for sequence analysis is described. The mass spectrometer is equipped with a matrix-assisted laser desorption/ionization (MALDI) source that operates at elevated pressure, 0.01-1 Torr. Ion fragmentation is controlled by varying the pressure of the buffer gas, the laser energy, the voltage difference between the MALDI target and the adjacent sampling cone, and between the cone and the quadrupole ion guide. The peptides were analyzed under optimal ionization conditions to obtain their molecular mass, and under conditions that promote ion dissociation via metastable decomposition or collision-induced dissociation (CID). The fragmentation spectra were used to obtain sequence information. Ion dissociation was promoted via three configurations of the ionization parameters. All methods yielded sequencing-grade b- and y-type ions. Two binary mixtures of peptides were used to demonstrate that: (1) external calibration provides a standard deviation (sigma) of 4 ppm with a mode of 9 ppm; and (2) that peptides with molecular masses that differ by a factor of two may be independently fragmented by appropriately choosing the CID energy and the low-mass cut-off. Analyses of tryptic digests employed liquid chromatography (LC), deposition of the eluant on a target, and finally MALDI-TOF mass spectrometry. The mass fingerprint and the (partial) sequence of the tryptic peptides were matched to their precursor protein via database searches. SN - 0951-4198 UR - https://www.unboundmedicine.com/medline/citation/15593255/Applications_of_a_matrix_assisted_laser_desorption/ionization_orthogonal_time_of_flight_mass_spectrometer__l__Metastable_decay_and_collision_induced_dissociation_for_sequencing_peptides_ L2 - https://doi.org/10.1002/rcm.1775 DB - PRIME DP - Unbound Medicine ER -