Methionine Sulfoxide Formation by Cigarette Smoke is Associated with the Degradation of Plasma Proteins
| Autor: | Abdullah Qassab, Rohana Liyanage, Wesley E. Stites |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Gel electrophoresis
Methionine Chromatography Chemistry Methionine sulfoxide Biophysics Protein turnover Urine Trypsin Human serum albumin Blood proteins respiratory tract diseases body regions chemistry.chemical_compound Biochemistry embryonic structures behavior and behavior mechanisms medicine medicine.drug |
| Zdroj: | Biophysical Journal. 112:48a-49a |
| ISSN: | 0006-3495 |
| DOI: | 10.1016/j.bpj.2016.11.301 |
| Popis: | Background: High level of protein carbonyls have been found in plasma proteins in smokers compared to non-smokers. While oxidation may directly interfere with activity, the extent to which oxidation affects protein turnover is less clear.Objectives: To determine levels of oxidized serum proteins cleared in the urine of smokers and non-smokers with focus on methionine sulfoxide (MSO) formation in Human Serum Albumin (HSA) and to determine the effect of methionine oxidation on the turnover of HSA.Method: 100 mL of urine were obtained from smoker and non-smokers. proteins were concentrated by reducing the sample size to 1.5% of the original volume. 200 µL of the concentrate then were separated by SDS-page gel electrophoresis. The band with intact HSA was cut out and the remainder of the gel was cut into four different pieces. Gel sections then were digested with trypsin. Levels of MSO in the resulting peptides were assessed by LC-MS/MS and data analysis was performed using the Skyline software package.Results: A group comparison between non-smokers (control) and smokers showed a slight increase in the levels of MSO found in intact HSA of smokers relative to non-smokers. Regions of gels with proteins of lower mass than intact HSA showed that degraded fragments of HSA in urine of both smokers and non-smokers have higher levels of MSO than are found in intact HSA.Conclusions: HSA in smokers has statistically significant higher levels of MSO than HSA in non-smokers. However, the higher levels of oxidation in smokers are concentrated in partially degraded HSA. At the moment it is not possible to say unequivocally whether oxidized HSA is more likely to be cleaved and cleared, if cleaved protein is more likely to be oxidized before clearance, or both. |
| Databáze: | OpenAIRE |
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