Effective novel dissociation methods for intact protein: heat-assisted nozzle-skimmer collisionally induced dissociation and infrared multiphoton dissociation using a Fourier transform ion cyclotron resonance mass spectrometer equipped with a micrometal electrospray ionization emitter.
Heating of a nano-electrospray ionization (nanoESI) source can improve the dissociation efficiency of collisionally induced dissociation (CID) methods, such as nozzle-skimmer CID (NS-CID) and infrared multiphoton dissociation (IRMPD), for large biomolecule fragmentation. A metal nanoESI emitter was used due to its resistance to heating above 250 degrees C. This novel method for the dissociation of large biomolecular ions is termed "heat-assisted NS-CID" (HANS-CID) or "heat-assisted IRMPD" (HA-IRMPD). Multiple charged nonreduced protein ions (8.6 Da ubiquitin, 14 kDa lysozyme, and 67 kDa bovine serum albumin) were directly dissociated by HANS-CID and HA-IRMPD to effectively yield fragment ions that could be assigned. The fragment ions of ubiquitin by HANS-CID can be analyzed by tandem mass spectrometry (MS/MS) using sustained off-resonance irradiation CID (SORI-CID) and IRMPD. In addition, a native large protein, immunoglobulin G (IgG, 150 kDa), was efficiently dissociated by HA-IRMPD. The product ions that were obtained reflected the domain structure of IgG. However, these product ions of IgG and lysozyme were not dissociated by MS/MS using the same heating energetic methods such as IRMPD and SORI-CID.
Yamada
Naoyuki
N
Institute of Life Sciences, Ajinomoto Company Inc., Kawasaki-ku, Kawasaki-shi 210-8681, Japan. naoyuki_yamada@ajinomoto.com
Suzuki
Ei-Ichiro
E
Hirayama
Kazuo
K
D000818:Animals; D002417:Cattle; D006358:Hot Temperature; D007074:Immunoglobulin G / Q000737:chemistry / Q000528:radiation effects; D007259:Infrared Rays; D009113:Muramidase / Q000737:chemistry / Q000528:radiation effects; D036103:Nanotechnology / Q000379:methods; D017785:Photons; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000528:radiation effects; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*; D017550:Spectroscopy, Fourier Transform Infrared / Q000295:instrumentation / Q000379:methods; D025801:Ubiquitin / Q000737:chemistry / Q000528:radiation effects
D007074:Immunoglobulin G; D025801:Ubiquitin; D012710:Serum Albumin, Bovine; D009113:Muramidase