Re: High Sensitivity Protein Identification

Axel Ducret (axel_ducret@merck.com)
Fri, 23 Jan 1998 10:44:42 -0500


Ken,

As a grant's reviewer, I would seriously doubt of a project that would propose
to do MS or MS/MS protein characterization at the low amol level or below. In my

knowledge, low fmol level (this is about what you will be able to detect with
silver staining on gel and western blotting ...) is probably the best you will
achieve (1 fmol of a 20 kDa protein = 20 PICOgrams ...).Here is a small (and
certainly not complete !) list of representative litterature:

for MALDI-Tof:

Caprioli RM, Farmer TB, Gile J. (1997) Molecular imaging of biological samples:
localization of peptides and proteins
using MALDI-TOF MS. Anal. Chem. 69(23) 4751-4760

Jensen ON, Podtelejnikov AV, Mann M (1997) Identification of the components of
simple protein mixtures by
high-accuracy peptide mass mapping and database searching. Anal. Chem. 69(23)
4741-4750

Cordwell SJ, Basseal DJ, Humphery-Smith I. (1997) Proteome analysis of
Spiroplasma melliferum (A56) and protein characterisation across species
boundaries.Electrophoresis 18(8) 1335-1346

Qin J, Fenyo D, Zhao Y, Hall WW, Chao DM, Wilson CJ, Young RA, Chait BT (1997) A

strategy for rapid, high-confidence protein identification. Anal. Chem. 69 (19)
3995-4001

Pappin DJ (1997) Peptide mass fingerprinting using MALDI-TOF mass spectrometry.
Methods Mol. Biol. 64 165-173

For CE and LC-MS/MS and nanoES-MS/MS:

McCormack AL, Schieltz DM, Goode B, Yang S, Barnes G, Drubin D, Yates JR 3rd
(1997) Direct analysis and identification of proteins in mixtures by LC/MS/MS
and database searching at the low-femtomole level. Anal Chem 69(4) 767-776

Davis MT, Stahl DC, Hefta SA, Lee TD (1995) A microscale electrospray interface
for on-line, capillary liquid
chromatography/tandem mass spectrometry of complex peptide mixtures. Anal. Chem.

67(24) 4549-4556

Tong D, Moritz RL, Eddes JS, Reid GE, Rasmussen RK, Dorow DS, Simpson RJ (1997)
Fabrication of stable packed capillary reversed-phase columns for protein
structural analysis. J. Prot. Chem. 16(5) 425-431

Wilm M, Shevchenko A, Houthaeve T, Breit S, Schweigerer L, Fotsis T, Mann M
(1996) Femtomole sequencing of proteins from polyacrylamide gels by
nano-electrospray mass spectrometry. Nature 379(6564) 466-469

Morris HR, Paxton T, Panico M, McDowell R, Dell A (1997) A novel geometry mass
spectrometer, the Q-TOF, for
low-femtomole/attomole-range biopolymer sequencing. J. Prot. Chem. 16(5) 469-479

Figeys D, Ducret A, Yates JR 3rd, Aebersold R. (1996) Protein identification by
solid-phase microextraction capillary-zone electrophoresis microelectrospray
tandem mass spectrometry. Nature Biotech.14 1579-1583

Wheat TE, Lilley KA, Banks JF (1997) Capillary electrophoresis with electrospray

mass spectrometry detection
for low-molecular-mass compounds. J. Chromatogr. A 781(1-2) 99-105

Valaskovic GA, Kelleher NL, McLafferty FW (1996) Attomole protein
characterization by capillary electrophoresis-mass
spectrometry. Science 273(5279) 1199-1202

Figeys D, Ning Y, Aebersold R (1997) A microfabricated device for rapid protein
identification by microelectrospray ion trap mass spectrometry. Anal. Chem.
69(16) 3153-3160

Xue Q, Foret F, Dunayevskiy YM, Zavracky PM, McGruer NE, Karger BL (1997)
Multichannel microchip electrospray mass spectrometry. Anal. Chem. 69(3) 426-430

Hope that it helps

Axel Ducret

--------------------------------------------------------------------------------
-----------------------------------

Axel Ducret, Ph.D.
Senior Research Biologist
Merck-Frosst Canada Inc.
Dept. Biochemistry and Molecular Biology
P.O. Box 1005
Pointe-Claire-Dorval PQ H9R 4P8
Canada

tel. + (514) 428-3428
fax + (514) 428-4900