Chemical Modifications of Peptides and Proteins with Low Concentration Formaldehyde Studied by Mass Spectrometry
10.11895/j.issn.0253-3820.160139
- VernacularTitle:低浓度甲醛对多肽和蛋白化学修饰的质谱研究
- Author:
Zijian WANG
;
Jingbo YANG
;
Guangpu LI
;
Ningning SUN
;
Wanchun SUN
;
Qisheng PENG
;
Ning LIU
- Publication Type:Journal Article
- Keywords:
Formaldehyde;
Peptide;
Matrix protein;
Chemical modification;
Mass spectrometry
- From:
Chinese Journal of Analytical Chemistry
2016;(8):1193-1199
- CountryChina
- Language:Chinese
-
Abstract:
Abstract Formaldehyde has been widely employed to immobilize clinical tissue specimens, inactivate toxins and viruses in biomedical fields. Formaldehyde can react with active groups in bio-molecules such as proteins, resulting in protein cross-linking, inactivation, and immobilization. By using several standard peptides and tryptic peptides from matrix protein of influenza virus as experimental models, we studied the chemical modifications of peptides and proteins with formaldehyde by matrix-assisted laser desorption ionization time-of-flight mass spectrometry and nano-electrospray quadruple time-of-flight tandem mass spectrometry. The reaction between formaldehyde and peptides was performed under the same conditions as those during inactivation of virus (4℃, 0. 025% Formalin (V/V), 37% formaldehyde solution (w/w), and 72 h). The results indicated that under above conditions, formaldehyde could react with amino group of N-terminus of standard peptide to generate a methylol adduct, which was further condensed into an imine to generate+12 Da product. Besides, formaldehyde could react with side chain of two amino acids such as arginine and lysine, yielding +12 Da product respectively. The analysis of the reaction between formaldehyde and tryptic peptides from matrix protein of influenza virus showed that +24 Da products could be detected in most peptides due to combinational contribution from N-terminus of peptide (+12 Da ) and side chain of C-terminal arginine or lysine (+12 Da) . Moreover, a +36 Da product was detected for a peptide with miss-cut site. The results indicated that low-concentration formaldehyde primarily reacted with amino group on N-termini of peptides and proteins, as well as the side chains of arginine and lysine residues. The present study suggested an effective mass spectrometry-based method for analyzing the reaction between low-concentration formaldehyde and peptides and proteins, thus provided strategies for interpretation for the mass spectra of reaction products.
