On particle ionization/enrichment of multifunctional nanoprobes: washing/separation-free, acceleration and enrichment of microwave-assisted tryptic digestion of proteins via bare TiO2 nanoparticles in ESI-MS and comparing to MALDI-MS.
A simple, rapid, straightforward and washing/separation free of in-solution digestion method for microwave-assisted tryptic digestion of proteins (cytochrome c, lysozyme and myoglobin) using bare TiO(2) nanoparticles (NPs) prepared in aqueous solution to serve as multifunctional nanoprobes in electrospray ionization mass spectrometry (ESI-MS) was demonstrated. The current approach is termed as 'on particle ionization/enrichment (OPIE)' and it can be applied in ESI-MS, atmospheric pressure-matrix-assisted laser desorption/ionization mass spectrometry (AP-MALDI-MS) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The bare TiO(2) NPs can assist, accelerate and effectively enhance the digestion efficiency, sequence coverage and detection sensitivity of peptides for the microwave-assisted tryptic digestion of proteins in ESI-MS. The reason is attributed to the fact that proteins or partially digested proteins are easily attracted or concentrated onto the surface of TiO(2) NPs, resulting in higher efficiency of digestion reactions in the microwave experiments. Besides, the TiO(2) NPs could act as a microwave absorber to accelerate and enrich the protein fragments in a short period of time (40-60 s) from the microwave experiments in ESI-MS. Furthermore, the bare TiO(2) NPs prepared in aqueous solution exhibit high adsorption capability toward the protein fragments (peptides); thus, the OPIE approach for detecting the digested protein fragments via ESI and MALDI ionization could be achieved. The current technique is also a washing and separation-free technique for accelerating and enriching microwave-assisted tryptic digestion of proteins in the ESI-MS and MALDI-MS. It exhibits potential to be widely applied to biotechnology and proteome research in the near future.
Department of Chemistry, National Sun Yat-Sen University, 70, Lien-Hai Road, Kaohsiung 80424, Taiwan. hwu@faculty.nsysu.edu.tw
Hui-Fen
HF
Wu
Kavita
K
Agrawal
Kamlesh
K
Shrivas
Yi-Hsien
YH
Lee
D000818:Animals; D019187:Cadmium Compounds / Q000737:chemistry; D002417:Cattle; D002645:Chickens; D045304:Cytochromes c / Q000737:chemistry / Q000378:metabolism; D006358:Hot Temperature; D008872:Microwaves; D015335:Molecular Probes / Q000737:chemistry / Q000378:metabolism*; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D045663:Quantum Dots; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013440:Sulfides / Q000737:chemistry; D014025:Titanium / Q000737:chemistry*; D014357:Trypsin / Q000737:chemistry* / Q000378:metabolism
D019187:Cadmium Compounds; D015335:Molecular Probes; D010446:Peptide Fragments; D013440:Sulfides; C034939:cadmium sulfide; C009495:titanium dioxide; D045304:Cytochromes c; D014025:Titanium; D009113:Muramidase; D014357:Trypsin