PMID- 15144972
OWN - NLM
STAT- MEDLINE
DCOM- 20040715
LR  - 20181113
IS  - 1044-0305 (Print)
IS  - 1044-0305 (Linking)
VI  - 15
IP  - 6
DP  - 2004 Jun
TI  - Mass spectrometric detection of affinity purified crosslinked peptides.
PG  - 832-9
AB  - Chemical crosslinking of proteins combined with mass spectrometric analysis of 
      the tryptic digest of the products shows considerable promise as a tool for 
      interrogating structure and geometry of proteins and protein complexes. An 
      impediment to the use of this tool has been the difficulty of distinguishing 
      crosslinked peptide pairs from non-crosslinked peptides, and from the products of 
      side reactions. We describe the use of a commercially available biotinylated 
      crosslinking reagent, sulfo-SBED, that allows affinity-based enrichment of 
      crosslinked species. An intramolecular crosslink is prepared using the peptide 
      neurotensin as a model system. Matrix-assisted laser desorption/ionization 
      time-of-flight (MALDI-TOF) mass spectra show the predicted crosslinking product, 
      as well as several side products. Finally, we describe the optimized enrichment 
      of biotinylated species, and reduction of non-specific binding, for a batch-mode 
      affinity separation based on immobilized monomeric avidin.
FAU - Hurst, Gregory B
AU  - Hurst GB
AD  - Chemical Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 
      37831-6131, USA. hurstgb@ornl.gov
FAU - Lankford, Trish K
AU  - Lankford TK
FAU - Kennel, Stephen J
AU  - Kennel SJ
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - J Am Soc Mass Spectrom
JT  - Journal of the American Society for Mass Spectrometry
JID - 9010412
RN  - 0 (Cross-Linking Reagents)
RN  - 0 (Peptides)
RN  - 1405-69-2 (Avidin)
SB  - IM
MH  - Animals
MH  - Avidin/metabolism
MH  - Cattle
MH  - Chromatography, Affinity
MH  - Cross-Linking Reagents/*chemistry
MH  - Peptides/*analysis/*chemistry/isolation & purification/metabolism
MH  - Sequence Analysis, Protein
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/*methods
EDAT- 2004/05/18 05:00
MHDA- 2004/07/16 05:00
CRDT- 2004/05/18 05:00
PHST- 2003/06/27 00:00 [received]
PHST- 2004/02/13 00:00 [revised]
PHST- 2004/02/13 00:00 [accepted]
PHST- 2004/05/18 05:00 [pubmed]
PHST- 2004/07/16 05:00 [medline]
PHST- 2004/05/18 05:00 [entrez]
AID - S1044030504001540 [pii]
AID - 10.1016/j.jasms.2004.02.008 [doi]
PST - ppublish
SO  - J Am Soc Mass Spectrom. 2004 Jun;15(6):832-9. doi: 10.1016/j.jasms.2004.02.008.