Variations in platelet protein associated with arterial thrombosis.
BACKGROUND
Hyperactivity of platelets has been associated with thrombotic episodes by molecular mechanisms not yet elucidated. The present work aimed at identifying whether the platelet protein content from patients who had suffered an arterial thrombosis episode differed from that of platelets obtained from normal healthy donors.


METHODS
Differential platelet protein profiles were determined by 2-dimensional (2-D) gel electrophoresis and Western blot analysis of total platelet lysates. Identification of differentially expressed proteins was carried out by mass spectrometry (MALDI-TOF).


RESULTS
We found a decreased platelet content of three protein spots in patients of arterial thrombosis: integrin linked kinase (ILK), fructose bisphosphate aldolase (aldolase) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) whereas the content of four other protein spots was increased: actin binding protein, coronine like (p57), non-muscle myosin heavy chain (NMMHC-A), pyruvate kinase M2 isoenzyme (PK) and phosphoglycerate kinase (PGK). The variations in ILK, GAPDH and PK were validated by Western blot analysis. The proteins showing a decreased platelet content in arterial thrombosis patients are associated with the cytoskeletal insoluble fraction and the detected increase in some proteins seems to be due to the generation of peptides caused by a limited proteolysis. Differences in the protein profiles of circulating platelets from arterial thrombosis were maintained months after the acute thrombotic event and disappear in the long term.


CONCLUSIONS
The observed variations in some platelet proteins suggest the existence of a perturbation in the cytoskeletal organization and increased proteolysis, both indicative of a platelet pro-active state, persistent after the thrombotic event.
Arias-Salgado
Elena G
EG
Department of Cellular and Molecular Physiopathology, Centre of Biological Research, CSIC, Madrid, Spain.
Larrucea
Susana
S
Butta
Nora
N
Fernández
Darío
D
García-Muñoz
Soledad
S
Parrilla
Roberto
R
Ayuso
Matilde S
MS
D000328:Adult; D001158:Arteries; D001792:Blood Platelets / Q000378:metabolism*; D001798:Blood Proteins / Q000378:metabolism*; D016022:Case-Control Studies; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D005634:Fructose-Bisphosphate Aldolase / Q000097:blood; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000097:blood; D006801:Humans; D008297:Male; D008875:Middle Aged; D017346:Protein Serine-Threonine Kinases / Q000097:blood; D011770:Pyruvate Kinase / Q000097:blood; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D020521:Stroke / Q000097:blood / Q000209:etiology; D013927:Thrombosis / Q000097:blood* / Q000209:etiology; D000099200:Scaffold Protein ILK
D001798:Blood Proteins; D005634:Fructose-Bisphosphate Aldolase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D017346:Protein Serine-Threonine Kinases; D011770:Pyruvate Kinase; D000099200:Scaffold Protein ILK