Identification of Plasma Protein Markers of Oxidation in Obese Men

Research output: Contribution to conferencePoster

Abstract

Aim: Pyrrole modifications to proteins are a sign of oxidative damage, they are not present in newly-formed proteins but occur when lipid oxidation products or sugars react non-enzymicly with free amino groups. Studying such modifications will enable the development of novel biomarkers of oxidative stress. We recently published methodology to detect and identify pyrrole-containing proteins by combining biotin-labelled Ehrlich’s reagent (ER-B) to specifically target protein bound pyrroles, 2D electrophoresis and LC/MS/MS. We hypothesize that plasma from obese subjects should contain pyrrolised proteins as the obese have increased oxidative stress.
Method: Our objective in this study was to use the technique described above to investigate the profile of these modified proteins in plasma samples from two groups of male age-matched volunteers, obese (BMI > 30) matched for adiposity (waist circumference) and lean (BMI 22-25) (n=4 per group).
Results: There was a discernable difference in the pattern of pyrrole-containing proteins between the two groups. All lean-plasma samples contained a small number of pyrrolised-proteins (up to a maximum of 4 proteins); while all obese-plasma samples had approximately four times as many different pyrrole-containing proteins. This demonstrates that this methodology has the sensitivity and specificity required to detect and identify proteins modified by oxidation in blood plasma from human subjects.
Original languageEnglish
Publication statusPublished - Apr 2011
Eventbiochemical society focussed meeting - Durham, United Kingdom
Duration: 20 Mar 201122 Mar 2011

Conference

Conferencebiochemical society focussed meeting
CountryUnited Kingdom
CityDurham
Period20/03/1122/03/11

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Blood Proteins
Pyrroles
Proteins
Oxidative Stress
Adiposity
Waist Circumference
Biotin
Leptin
Electrophoresis
Volunteers
Age Groups
Biomarkers
Lipids
Sensitivity and Specificity

Cite this

Campbell, F. M., Robins, S., & Hoggard, N. (2011). Identification of Plasma Protein Markers of Oxidation in Obese Men. Poster session presented at biochemical society focussed meeting, Durham, United Kingdom.

Identification of Plasma Protein Markers of Oxidation in Obese Men. / Campbell, Fiona Margaret; Robins, Simon; Hoggard, Nigel.

2011. Poster session presented at biochemical society focussed meeting, Durham, United Kingdom.

Research output: Contribution to conferencePoster

Campbell, FM, Robins, S & Hoggard, N 2011, 'Identification of Plasma Protein Markers of Oxidation in Obese Men' biochemical society focussed meeting, Durham, United Kingdom, 20/03/11 - 22/03/11, .
Campbell FM, Robins S, Hoggard N. Identification of Plasma Protein Markers of Oxidation in Obese Men. 2011. Poster session presented at biochemical society focussed meeting, Durham, United Kingdom.
Campbell, Fiona Margaret ; Robins, Simon ; Hoggard, Nigel. / Identification of Plasma Protein Markers of Oxidation in Obese Men. Poster session presented at biochemical society focussed meeting, Durham, United Kingdom.
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AU - Robins, Simon

AU - Hoggard, Nigel

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N2 - Aim: Pyrrole modifications to proteins are a sign of oxidative damage, they are not present in newly-formed proteins but occur when lipid oxidation products or sugars react non-enzymicly with free amino groups. Studying such modifications will enable the development of novel biomarkers of oxidative stress. We recently published methodology to detect and identify pyrrole-containing proteins by combining biotin-labelled Ehrlich’s reagent (ER-B) to specifically target protein bound pyrroles, 2D electrophoresis and LC/MS/MS. We hypothesize that plasma from obese subjects should contain pyrrolised proteins as the obese have increased oxidative stress. Method: Our objective in this study was to use the technique described above to investigate the profile of these modified proteins in plasma samples from two groups of male age-matched volunteers, obese (BMI > 30) matched for adiposity (waist circumference) and lean (BMI 22-25) (n=4 per group). Results: There was a discernable difference in the pattern of pyrrole-containing proteins between the two groups. All lean-plasma samples contained a small number of pyrrolised-proteins (up to a maximum of 4 proteins); while all obese-plasma samples had approximately four times as many different pyrrole-containing proteins. This demonstrates that this methodology has the sensitivity and specificity required to detect and identify proteins modified by oxidation in blood plasma from human subjects.

AB - Aim: Pyrrole modifications to proteins are a sign of oxidative damage, they are not present in newly-formed proteins but occur when lipid oxidation products or sugars react non-enzymicly with free amino groups. Studying such modifications will enable the development of novel biomarkers of oxidative stress. We recently published methodology to detect and identify pyrrole-containing proteins by combining biotin-labelled Ehrlich’s reagent (ER-B) to specifically target protein bound pyrroles, 2D electrophoresis and LC/MS/MS. We hypothesize that plasma from obese subjects should contain pyrrolised proteins as the obese have increased oxidative stress. Method: Our objective in this study was to use the technique described above to investigate the profile of these modified proteins in plasma samples from two groups of male age-matched volunteers, obese (BMI > 30) matched for adiposity (waist circumference) and lean (BMI 22-25) (n=4 per group). Results: There was a discernable difference in the pattern of pyrrole-containing proteins between the two groups. All lean-plasma samples contained a small number of pyrrolised-proteins (up to a maximum of 4 proteins); while all obese-plasma samples had approximately four times as many different pyrrole-containing proteins. This demonstrates that this methodology has the sensitivity and specificity required to detect and identify proteins modified by oxidation in blood plasma from human subjects.

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