Ice as a matrix for IR-matrix-assisted laser desorption/ionization: mass spectra from a protein single crystal.
Lasers emitting in the ultraviolet wavelength range of 260-360 nm are almost exclusively used for matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) of macromolecules. Reports about the use of lasers emitting in the infrared first appeared in 1990/1991. In contrast to MALDI in the ultraviolet, a very limited number of reports on IR-MALDI have since been published. Several matrices have been identified for infrared MALDI yielding spectra of a quality comparable to those obtained in the ultraviolet. Water (ice) was recognized early as a potential matrix because of its strong O-H stretching mode near 3 microm. Interest in water as matrix derives primarily from the fact that it is the major constituent of most biological tissues. If functional as matrix, it might allow the in situ analysis of macromolecular constituents in frozen cell sections without extraction or exchanging the water. We present results that show that IR-MALDI of lyophilized proteins, air dried protein solutions, or protein crystals up to a molecular mass of 30 kDa is possible without the addition of any separate matrix. Samples must be frozen to retain a sufficient fraction of the water of hydration in the vacuum. The limited current sensitivity, requiring at least 10 pmol of protein for a successful analysis needs to be further improved.
Berkenkamp
S
S
Institute for Medical Physics, University of Münster, Germany.
Karas
M
M
Hillenkamp
F
F
D000818:Animals; D002417:Cattle; D002645:Chickens; D003460:Crystallization; D003574:Cytochrome c Group / Q000737:chemistry; D005260:Female; D007053:Ice; D007328:Insulin / Q000737:chemistry; D009113:Muramidase / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013055:Spectrophotometry, Infrared / Q000379:methods*; D014452:Ubiquitins / Q000737:chemistry
D003574:Cytochrome c Group; D007053:Ice; D007328:Insulin; D011506:Proteins; D014452:Ubiquitins; D012710:Serum Albumin, Bovine; D009113:Muramidase