PMID- 9784743
OWN - NLM
STAT- MEDLINE
DCOM- 19981118
LR  - 20190717
IS  - 0003-2700 (Print)
IS  - 0003-2700 (Linking)
VI  - 70
IP  - 19
DP  - 1998 Oct 1
TI  - Measurement of amide hydrogen exchange by MALDI-TOF mass spectrometry.
PG  - 3987-95
AB  - Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass 
      spectrometry (MS) was used to determine amide proton/deuteron (H/D) exchange 
      rates. The method has broad application to the study of protein conformation and 
      folding and to the study of protein-ligand interactions and requires no 
      modifications of the instrument. Amide protons were allowed to exchange with 
      deuterons in buffered D2O at room temperature, pD 7.25. Exchanged deuterons were 
      "frozen" in the exchanged state by quenching at pH 2.5, 0 degree C and analyzed 
      by MALDI-TOF MS. The matrix mixture consisted of 5 mg/mL 
      alpha-cyano-4-hydroxycinnamic acid, acetonitrile, ethanol, and 0.1% TFA. The 
      matrix was adjusted to pH 2.5, and the chilled MALDI target was rapidly dried. 
      Deuteration of amide protons on cyclic AMP-dependent protein kinase was measured 
      after short times of incubation in deuterium by pepsin protein digestion and 
      MALDI-TOF MS analysis. The unseparated peptic digest was analyzed in a single 
      spectrum of the mixture. From five spectra, H/D exchange rates were determined 
      for some 40 peptides covering 65% of the protein sequence.
FAU - Mandell, J G
AU  - Mandell JG
AD  - Department of Chemistry and Biochemistry, University of California, San Diego, La 
      Jolla 92093-0601, USA. ekomives@ucsd.edu
FAU - Falick, A M
AU  - Falick AM
FAU - Komives, E A
AU  - Komives EA
LA  - eng
GR  - HL47463/HL/NHLBI NIH HHS/United States
GR  - T32 CA09523/CA/NCI NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Amides)
RN  - 0 (Peptide Fragments)
RN  - 7YNJ3PO35Z (Hydrogen)
RN  - 9041-90-1 (Angiotensin I)
RN  - AR09D82C7G (Deuterium)
RN  - EC 2.7.11.11 (Cyclic AMP-Dependent Protein Kinases)
RN  - EC 3.4.23.1 (Pepsin A)
SB  - IM
MH  - Amides/*analysis
MH  - Amino Acid Sequence
MH  - Angiotensin I/chemistry
MH  - Cyclic AMP-Dependent Protein Kinases/*chemistry
MH  - Deuterium/chemistry
MH  - Hydrogen/chemistry
MH  - Molecular Sequence Data
MH  - Pepsin A/metabolism
MH  - Peptide Fragments/*analysis
MH  - *Protein Conformation
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/*methods
EDAT- 1998/10/24 00:00
MHDA- 1998/10/24 00:01
CRDT- 1998/10/24 00:00
PHST- 1998/10/24 00:00 [pubmed]
PHST- 1998/10/24 00:01 [medline]
PHST- 1998/10/24 00:00 [entrez]
AID - 10.1021/ac980553g [doi]
PST - ppublish
SO  - Anal Chem. 1998 Oct 1;70(19):3987-95. doi: 10.1021/ac980553g.