Preparing protein microarrays by soft-landing of mass-selected ions.
Intact, multiply protonated proteins of particular mass and charge were selected from ionized protein mixtures and gently landed at different positions on a surface to form a microarray. An array of cytochrome c, lysozyme, insulin, and apomyoglobin was generated, and the deposited proteins showed electrospray ionization mass spectra that matched those of the authentic compounds. Deposited lysozyme and trypsin retained their biological activity. Multiply charged ions of protein kinase A catalytic subunit and hexokinase were also soft-landed into glycerol-based liquid surfaces. These soft-landed kinases phosphorylated LRRASLG oligopeptide and D-fructose, respectively.
Ouyang
Zheng
Z
Department of Chemistry, Purdue University, West Lafayette, IN 47907, USA.
Takáts
Zoltán
Z
Blake
Thomas A
TA
Gologan
Bogdan
B
Guymon
Andy J
AJ
Wiseman
Justin M
JM
Oliver
Justin C
JC
Davisson
V Jo
VJ
Cooks
R Graham
RG
D001059:Apoproteins / Q000737:chemistry; D020134:Catalytic Domain; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism; D003574:Cytochrome c Group / Q000737:chemistry / Q000378:metabolism; D006593:Hexokinase / Q000378:metabolism; D007328:Insulin / Q000737:chemistry; D007477:Ions; D013058:Mass Spectrometry; D009113:Muramidase / Q000378:metabolism; D009211:Myoglobin / Q000737:chemistry; D009496:Neurotensin / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism; D010766:Phosphorylation; D040081:Protein Array Analysis; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism
D001059:Apoproteins; D003574:Cytochrome c Group; D007328:Insulin; D007477:Ions; D009211:Myoglobin; D009842:Oligopeptides; D009844:Oligosaccharides; D011506:Proteins; C010857:apomyoglobin; C056529:N-acetylchitohexaose; D009496:Neurotensin; C037642:kemptide; D006593:Hexokinase; D017868:Cyclic AMP-Dependent Protein Kinases; D009113:Muramidase; D014357:Trypsin