Proteomic identification of differentially expressed proteins in aortic wall of patients with ruptured and nonruptured abdominal aortic aneurysms.
OBJECTIVE
To compare the basic proteomic composition of abdominal aortic aneurysm (AAA) wall tissue in patients with nonruptured and ruptured aneurysms.


METHODS
A proteomic approach with two-dimensional gel electrophoresis (2D-PAGE) and mass spectrometry (MS) was used to identify differentially expressed proteins in AAA tissue from nine patients with nonruptured and eight patients with ruptured AAA. Computerized image analysis was used to detect protein spots. Differentially expressed protein spots were in-gel digested and identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Western blot analysis was used to confirm differential expression.


RESULTS
Seven differentially expressed proteins were detected among 745 protein spots, selecting spots whose average relative volumes differed more than twofold between the nonruptured and the ruptured group. Four protein spots were up-regulated in the ruptured group, and three were down-regulated. Five of the spots were identified. Among the upregulated spots, No. 605 was identified as peroxiredoxin-2. The up-regulation was confirmed by Western blotting. No. 381 was identified as an actin fragment. Two spots, Nos. 719 and 499, could not be identified. Among the down-regulated protein spots, No. 130 contained two peptides; one reliably determined peptide, FEDGVLDPDYPR, is found in vitronectin. Another peptide, QIDNPDYK, was borderline significant and found in calreticulin. The down-regulation of vitronectin was confirmed by Western blotting. Spot Nos. 193 and 199 both contained peptides from albumin with actin also present in No. 199.


CONCLUSIONS
The identified proteins suggest that the aortic wall of ruptured aneurysms responds to a stressful condition and that proteolytic degradation of the cytoskeleton and connective tissue may be part of the response.
Urbonavicius
Sigitas
S
Department of Vascular Surgery, Vascular Research Unit, Viborg Hospital, Viborg, Denmark. sigitas.urbonavicius@sygehusviborg.dk
Lindholt
Jes S
JS
Vorum
Henrik
H
Urbonaviciene
Grazina
G
Henneberg
Eskild W
EW
Honoré
Bent
B
D000199:Actins / Q000032:analysis; D000368:Aged; D000369:Aged, 80 and over; D000418:Albumins / Q000032:analysis; D000595:Amino Acid Sequence; D001012:Aorta, Abdominal / Q000737:chemistry*; D017544:Aortic Aneurysm, Abdominal / Q000378:metabolism*; D001019:Aortic Rupture / Q000378:metabolism*; D015415:Biomarkers / Q000032:analysis; D015153:Blotting, Western; D037282:Calreticulin / Q000032:analysis; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D054464:Peroxiredoxins / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D012815:Signal Processing, Computer-Assisted; D053719:Tandem Mass Spectrometry; D019096:Vitronectin / Q000032:analysis
D000199:Actins; D000418:Albumins; D015415:Biomarkers; D037282:Calreticulin; D010446:Peptide Fragments; D011506:Proteins; D019096:Vitronectin; C086065:PRDX2 protein, human; D054464:Peroxiredoxins