Investigating the reaction of a novel silica capillary coating compound with proteins/peptides by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry.
Quaternized piperazine ((N-methyl-N-omega-iodobutyl-N'-methyl)piperazine; QPzl) is a novel compound described as an ideal coating material for the silica capillaries that are commonly used for capillary zone electrophoresis. In the course of such analysis, contact between such coatings and biomolecules may result in certain modifications of the latter. To gain specific information on such potential modifications, solutions at pH 10.0 containing both QPzl and standard proteins/peptides were incubated for various periods and examined by matrix-assisted laser desorption/ionisation mass spectrometry. The reduction of the S-S bridges, denaturation in 8 M urea, the isoelectric point of the protein and the duration of the incubation had a profound influence on the investigated reaction. Analysis in reflectron mode and post source decay identified Cys as the likely site of interaction. The implications of the present measurements for proteome analysis using capillary and gel electrophoresis are discussed.
Galvani
M
M
GlaxoWellcome Medicines Research Center, via Fleming 4, 37135 Verona, Italy.
Hamdan
M
M
Righetti
P G
PG
Gelfi
C
C
Sebastiano
R
R
Citterio
A
A
D000818:Animals; D002417:Cattle; D006863:Hydrogen-Ion Concentration; D010879:Piperazines / Q000737:chemistry*; D012259:Ribonuclease, Pancreatic / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013482:Superoxide Dismutase / Q000737:chemistry
D010879:Piperazines; D012822:Silicon Dioxide; D013482:Superoxide Dismutase; D012259:Ribonuclease, Pancreatic