Proteomic Investigation on Grp94-IgG Complexes Circulating in Plasma of Type 1 Diabetic Subjects
The glucose-regulated protein94 (Grp94) has been found in complexes with IgG in plasma of Type 1 (T1) diabetic subjects; however, the pathogenetic meaning of Grp94-IgG complexes has not yet been elucidated. To shed light on the nature and structure of these complexes in vivo, we conducted a proteomi...
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| Format: | Article |
| Language: | English |
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Wiley
2015-01-01
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| Series: | Journal of Diabetes Research |
| Online Access: | http://dx.doi.org/10.1155/2015/815839 |
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| author | Antonella Roveri Mattia Zaccarin Andrea Pagetta Elisa Tramentozzi Paola Finotti |
| author_facet | Antonella Roveri Mattia Zaccarin Andrea Pagetta Elisa Tramentozzi Paola Finotti |
| author_sort | Antonella Roveri |
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| description | The glucose-regulated protein94 (Grp94) has been found in complexes with IgG in plasma of Type 1 (T1) diabetic subjects; however, the pathogenetic meaning of Grp94-IgG complexes has not yet been elucidated. To shed light on the nature and structure of these complexes in vivo, we conducted a proteomic analysis on plasma of both T1 diabetic subjects and healthy control subjects. IgG purified from plasma was submitted to 2D PAGE followed by Western blotting and mass analysis. Grp94 was detected in plasma of all diabetic but not control subjects and found linked with its N-terminus to the IgG heavy chain. Mass analysis of heavy chain of IgG that binds Grp94 also in vitro, forming stable complexes with characteristics similar to those of native ones, permitted identifying CH2 and CH3 regions as those involved in binding Grp94. At the electron microscopy, IgG from diabetic plasma appeared as fibrils of various lengthes and dimensions, suggestive of elevated aggregating tendency conferred to IgG by Grp94. The nonimmune nature of complexes turned out to be responsible for the particular stability and structure adopted by complexes in plasma of diabetic subjects. Results are of relevance to understanding the pathogenetic mechanisms underlying diabetes and its complications. |
| format | Article |
| id | doaj-art-a4ee9bf8d95847c2b65ea03990126945 |
| institution | Kabale University |
| issn | 2314-6745 2314-6753 |
| language | English |
| publishDate | 2015-01-01 |
| publisher | Wiley |
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| series | Journal of Diabetes Research |
| spelling | doaj-art-a4ee9bf8d95847c2b65ea039901269452025-08-20T03:38:13ZengWileyJournal of Diabetes Research2314-67452314-67532015-01-01201510.1155/2015/815839815839Proteomic Investigation on Grp94-IgG Complexes Circulating in Plasma of Type 1 Diabetic SubjectsAntonella Roveri0Mattia Zaccarin1Andrea Pagetta2Elisa Tramentozzi3Paola Finotti4Department of Molecular Medicine, Section of Biological Chemistry, University of Padua, Via G. Colombo 3, 35131 Padua, ItalyDepartment of Molecular Medicine, Section of Biological Chemistry, University of Padua, Via G. Colombo 3, 35131 Padua, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padua, Largo E. Meneghetti 2, 35131 Padua, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padua, Largo E. Meneghetti 2, 35131 Padua, ItalyDepartment of Pharmaceutical and Pharmacological Sciences, University of Padua, Largo E. Meneghetti 2, 35131 Padua, ItalyThe glucose-regulated protein94 (Grp94) has been found in complexes with IgG in plasma of Type 1 (T1) diabetic subjects; however, the pathogenetic meaning of Grp94-IgG complexes has not yet been elucidated. To shed light on the nature and structure of these complexes in vivo, we conducted a proteomic analysis on plasma of both T1 diabetic subjects and healthy control subjects. IgG purified from plasma was submitted to 2D PAGE followed by Western blotting and mass analysis. Grp94 was detected in plasma of all diabetic but not control subjects and found linked with its N-terminus to the IgG heavy chain. Mass analysis of heavy chain of IgG that binds Grp94 also in vitro, forming stable complexes with characteristics similar to those of native ones, permitted identifying CH2 and CH3 regions as those involved in binding Grp94. At the electron microscopy, IgG from diabetic plasma appeared as fibrils of various lengthes and dimensions, suggestive of elevated aggregating tendency conferred to IgG by Grp94. The nonimmune nature of complexes turned out to be responsible for the particular stability and structure adopted by complexes in plasma of diabetic subjects. Results are of relevance to understanding the pathogenetic mechanisms underlying diabetes and its complications.http://dx.doi.org/10.1155/2015/815839 |
| spellingShingle | Antonella Roveri Mattia Zaccarin Andrea Pagetta Elisa Tramentozzi Paola Finotti Proteomic Investigation on Grp94-IgG Complexes Circulating in Plasma of Type 1 Diabetic Subjects Journal of Diabetes Research |
| title | Proteomic Investigation on Grp94-IgG Complexes Circulating in Plasma of Type 1 Diabetic Subjects |
| title_full | Proteomic Investigation on Grp94-IgG Complexes Circulating in Plasma of Type 1 Diabetic Subjects |
| title_fullStr | Proteomic Investigation on Grp94-IgG Complexes Circulating in Plasma of Type 1 Diabetic Subjects |
| title_full_unstemmed | Proteomic Investigation on Grp94-IgG Complexes Circulating in Plasma of Type 1 Diabetic Subjects |
| title_short | Proteomic Investigation on Grp94-IgG Complexes Circulating in Plasma of Type 1 Diabetic Subjects |
| title_sort | proteomic investigation on grp94 igg complexes circulating in plasma of type 1 diabetic subjects |
| url | http://dx.doi.org/10.1155/2015/815839 |
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