Attomole quantitation of protein separations with accelerator mass spectrometry.
Quantification of specific proteins depends on separation by chromatography or electrophoresis followed by chemical detection schemes such as staining and fluorophore adhesion. Chemical exchange of short-lived isotopes, particularly sulfur, is also prevalent despite the inconveniences of counting radioactivity. Physical methods based on isotopic and elemental analyses offer highly sensitive protein quantitation that has linear response over wide dynamic ranges and is independent of protein conformation. Accelerator mass spectrometry quantifies long-lived isotopes such as 14C to subattomole sensitivity. We quantified protein interactions with small molecules such as toxins, vitamins, and natural biochemicals at precisions of 1-5%. Micro-proton-induced X-ray emission quantifies elemental abundances in separated metalloprotein samples to nanogram amounts and is capable of quantifying phopsphorylated loci in gels. Accelerator-based quantitation is a possible tool for quantifying the genome translation into proteome.
Vogel
J S
JS
Center for Accelerator Mass Spectrometry, Lawrence Livermore National Laboratory, CA 94551, USA. jsvogel@llnl.gov
Grant
P G
PG
Buchholz
B A
BA
Dingley
K
K
Turteltaub
K W
KW
D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D002250:Carbon Radioisotopes; D002851:Chromatography, High Pressure Liquid; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D066298:In Vitro Techniques; D013058:Mass Spectrometry / Q000379:methods*; D008667:Metalloproteins / Q000032:analysis / Q000302:isolation & purification; D010599:Pharmacokinetics; D011485:Protein Binding; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D020543:Proteome / Q000032:analysis / Q000302:isolation & purification; D013052:Spectrometry, X-Ray Emission / Q000379:methods
D000596:Amino Acids; D002250:Carbon Radioisotopes; D008667:Metalloproteins; D011506:Proteins; D020543:Proteome