Ero1alpha is expressed on blood platelets in association with protein-disulfide isomerase and contributes to redox-controlled remodeling of alphaIIbbeta3.
Recent evidence supports a role of protein-disulfide isomerase (PDI) in redox-controlled remodeling of the exofacial domains of α(IIb)β(3) in blood platelets. The aim of this study was to explain whether Ero1α can be responsible for extracellular reoxidation of the PDI active site. We showed that Ero1α can be found on platelets and is rapidly recruited to the cell surface in response to platelet agonists. It is physically associated with PDI and α(IIb)β(3), as suggested by colocalization analysis in confocal microscopy and confirmed by immunoprecipitation experiments. Apart from monomeric oxidized Ero1α, anti-α(IIb)β(3) immunoprecipitates showed the presence of several Ero1α-positive bands that corresponded to the complexes α(IIb)β(3)-PDI-Ero1α, PDI-Ero1α, and Ero1α-Ero1α dimers. It binds more efficiently to the activated α(IIb)β(3) conformer, and its interaction is inhibited by RGD peptides. Ero1α appears to be involved in the regulation of α(IIb)β(3) receptor activity because of the following: (a) blocking the cell surface Ero1α by antibodies leads to a decrease in platelet aggregation in response to agonists and a decrease in fibrinogen and PAC-1 binding, and (b) transfection of MEG01 with Ero1α increases α(IIb)β(3) receptor activity, as indicated by increased binding of fibrinogen.
Swiatkowska
Maria
M
Department of Molecular and Medical Biophysics, Medical University of Lodz, 92-215 Lodz, Poland.
Padula
Gianluca
G
Michalec
Lidia
L
Stasiak
Marta
M
Skurzynski
Szymon
S
Cierniewski
Czeslaw S
CS
D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism; D001792:Blood Platelets / Q000378:metabolism*; D020134:Catalytic Domain; D005340:Fibrinogen / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism*; D009842:Oligopeptides / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D010974:Platelet Aggregation / Q000187:drug effects / Q000502:physiology*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D019704:Protein Disulfide-Isomerases / Q000235:genetics / Q000378:metabolism*
D000951:Antigens, Neoplasm; D008562:Membrane Glycoproteins; D046912:Multiprotein Complexes; D009842:Oligopeptides; C505096:PCA-1 antigen, human; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; C047981:arginyl-glycyl-aspartic acid; D005340:Fibrinogen; C405327:ERO1A protein, human; D010088:Oxidoreductases; D019704:Protein Disulfide-Isomerases