Infrared-assisted tryptic proteolysis for peptide mapping.
In this report, infrared (IR) radiation was employed to enhance the efficiency of tryptic proteolysis for peptide mapping. Protein solutions containing trypsin in sealed transparent Eppendorf tubes were allowed to digest under an IR lamp at 37 degrees C. The feasibility and performance of the novel proteolysis approach were demonstrated by the digestion of BSA and myoglobin (MYO) and the digestion time was significantly reduced to 5 min. The obtained digests were identified by MALDI-TOF MS with the sequence coverages of 69% (BSA) and 90% (MYO) that were much better than those obtained by conventional in-solution tryptic digestion. The present IR-assisted proteolysis strategy is simple and efficient, offering great promise for high-throughput protein identification.
School of Pharmacy and Department of Chemistry, Fudan University, Shanghai, China.
Sheng
S
Wang
Luyan
L
Zhang
Pengyuan
P
Yang
Gang
G
Chen
D000818:Animals; D001639:Bicarbonates / Q000737:chemistry; D002021:Buffers; D002417:Cattle; D002645:Chickens; D005240:Feasibility Studies; D006736:Horses; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis / Q000187:drug effects / Q000528:radiation effects; D007259:Infrared Rays; D009113:Muramidase / Q000737:chemistry; D009211:Myoglobin / Q000737:chemistry; D010447:Peptide Hydrolases / Q000378:metabolism*; D010449:Peptide Mapping / Q000295:instrumentation* / Q000379:methods*; D015203:Reproducibility of Results; D012710:Serum Albumin, Bovine / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D017550:Spectroscopy, Fourier Transform Infrared; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism*; D014732:Vibration
D001639:Bicarbonates; D002021:Buffers; D009211:Myoglobin; D012710:Serum Albumin, Bovine; D009113:Muramidase; D010447:Peptide Hydrolases; D014357:Trypsin