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Transthyretin Amyloidosis: Chaperone Concentration Changes and Increased Proteolysis in the Pathway to Disease.

da Costa G, Ribeiro-Silva C, Ribeiro R, Gilberto S, Gomes RA, Ferreira A, Mateus É, Barroso E, Coelho AV, Freire AP, Cordeiro C - PLoS ONE (2015)

Bottom Line: Transthyretin amyloidosis is a conformational pathology characterized by the extracellular formation of amyloid deposits and the progressive impairment of the peripheral nervous system.Point mutations in this tetrameric plasma protein decrease its stability and are linked to disease onset and progression.We discovered, using a differential proteomics approach, that extracellular chaperones such as fibrinogen, clusterin, haptoglobin, alpha-1-anti-trypsin and 2-macroglobulin are overrepresented in transthyretin amyloidosis.

View Article: PubMed Central - PubMed

Affiliation: Centro de Química e Bioquímica, FCUL, Campo Grande, Lisboa, Portugal.

ABSTRACT
Transthyretin amyloidosis is a conformational pathology characterized by the extracellular formation of amyloid deposits and the progressive impairment of the peripheral nervous system. Point mutations in this tetrameric plasma protein decrease its stability and are linked to disease onset and progression. Since non-mutated transthyretin also forms amyloid in systemic senile amyloidosis and some mutation bearers are asymptomatic throughout their lives, non-genetic factors must also be involved in transthyretin amyloidosis. We discovered, using a differential proteomics approach, that extracellular chaperones such as fibrinogen, clusterin, haptoglobin, alpha-1-anti-trypsin and 2-macroglobulin are overrepresented in transthyretin amyloidosis. Our data shows that a complex network of extracellular chaperones are over represented in human plasma and we speculate that they act synergistically to cope with amyloid prone proteins. Proteostasis may thus be as important as point mutations in transthyretin amyloidosis.

No MeSH data available.


Related in: MedlinePlus

Proteome analysis of plasma from ATTR individuals.A– 2D-PAGE analysis of plasma proteins. Labeled spots showa statistically significant variation (p<0.05) and a minimal fold variation of 1.5. These spots were excised, tryptic digested and proteins identified by MS/MS analysis. Average normalized volumes and protein identifications are presented in Table 1. B–Principle component analysis (PCA) of the 2D results. Each data point in the PCA represents the global expression values for all spots with a significant ANOVA value (p<0.05). A separation between the control and the ATTR individuals is clearly observed. C- 2D image analysis of four protein spots and normalized volumes, shown as examples. D-Over expression of western blot analysis of plasma from four control and four FAP individuals to detect TTR. E—Western blot analysis of a 2DE of serum from four control and four FAP individuals to detect TTR with super imposition of spots identified as TTR in 2DE.
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pone.0125392.g001: Proteome analysis of plasma from ATTR individuals.A– 2D-PAGE analysis of plasma proteins. Labeled spots showa statistically significant variation (p<0.05) and a minimal fold variation of 1.5. These spots were excised, tryptic digested and proteins identified by MS/MS analysis. Average normalized volumes and protein identifications are presented in Table 1. B–Principle component analysis (PCA) of the 2D results. Each data point in the PCA represents the global expression values for all spots with a significant ANOVA value (p<0.05). A separation between the control and the ATTR individuals is clearly observed. C- 2D image analysis of four protein spots and normalized volumes, shown as examples. D-Over expression of western blot analysis of plasma from four control and four FAP individuals to detect TTR. E—Western blot analysis of a 2DE of serum from four control and four FAP individuals to detect TTR with super imposition of spots identified as TTR in 2DE.

Mentions: Plasma proteins from controls and ATTR were separated by 2-D PAGE and analyzed with the SameSpots image analysis software. Fig 1A represents a standard gel image obtained by the use of the SameSpots analysis software, combining the Gaussian images of each gel. This latter was used as a reference master map for comparison of each 2-DE gel image from ATTR and controls, in the search of differentially expressed plasma proteins. The protein spot patterns were reproducible among individuals within a study group. Principal component analysis (PCA) revealed that control individuals show a marked difference from ATTR patients. We observed clusters of 3 replicates in each group, each cluster corresponding to one individual, with the three assays corresponding to the three experimental replicates performed (Fig 1B). Similarity between 2-D PAGE gels was very high, since 95% of spots were detected in 80% of gels from individual samples. An ANOVA analysis with a cutoff at 1.5 (p < 0.05) revealed 42 differentially expressed spots (Table 1) between ATTR and control individuals, as shown in Fig 1C. Table 1 contains informations of the identified proteins, including their accession number, matched peptides as well as theoretical and experimental molecular weight.


Transthyretin Amyloidosis: Chaperone Concentration Changes and Increased Proteolysis in the Pathway to Disease.

da Costa G, Ribeiro-Silva C, Ribeiro R, Gilberto S, Gomes RA, Ferreira A, Mateus É, Barroso E, Coelho AV, Freire AP, Cordeiro C - PLoS ONE (2015)

Proteome analysis of plasma from ATTR individuals.A– 2D-PAGE analysis of plasma proteins. Labeled spots showa statistically significant variation (p<0.05) and a minimal fold variation of 1.5. These spots were excised, tryptic digested and proteins identified by MS/MS analysis. Average normalized volumes and protein identifications are presented in Table 1. B–Principle component analysis (PCA) of the 2D results. Each data point in the PCA represents the global expression values for all spots with a significant ANOVA value (p<0.05). A separation between the control and the ATTR individuals is clearly observed. C- 2D image analysis of four protein spots and normalized volumes, shown as examples. D-Over expression of western blot analysis of plasma from four control and four FAP individuals to detect TTR. E—Western blot analysis of a 2DE of serum from four control and four FAP individuals to detect TTR with super imposition of spots identified as TTR in 2DE.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4492746&req=5

pone.0125392.g001: Proteome analysis of plasma from ATTR individuals.A– 2D-PAGE analysis of plasma proteins. Labeled spots showa statistically significant variation (p<0.05) and a minimal fold variation of 1.5. These spots were excised, tryptic digested and proteins identified by MS/MS analysis. Average normalized volumes and protein identifications are presented in Table 1. B–Principle component analysis (PCA) of the 2D results. Each data point in the PCA represents the global expression values for all spots with a significant ANOVA value (p<0.05). A separation between the control and the ATTR individuals is clearly observed. C- 2D image analysis of four protein spots and normalized volumes, shown as examples. D-Over expression of western blot analysis of plasma from four control and four FAP individuals to detect TTR. E—Western blot analysis of a 2DE of serum from four control and four FAP individuals to detect TTR with super imposition of spots identified as TTR in 2DE.
Mentions: Plasma proteins from controls and ATTR were separated by 2-D PAGE and analyzed with the SameSpots image analysis software. Fig 1A represents a standard gel image obtained by the use of the SameSpots analysis software, combining the Gaussian images of each gel. This latter was used as a reference master map for comparison of each 2-DE gel image from ATTR and controls, in the search of differentially expressed plasma proteins. The protein spot patterns were reproducible among individuals within a study group. Principal component analysis (PCA) revealed that control individuals show a marked difference from ATTR patients. We observed clusters of 3 replicates in each group, each cluster corresponding to one individual, with the three assays corresponding to the three experimental replicates performed (Fig 1B). Similarity between 2-D PAGE gels was very high, since 95% of spots were detected in 80% of gels from individual samples. An ANOVA analysis with a cutoff at 1.5 (p < 0.05) revealed 42 differentially expressed spots (Table 1) between ATTR and control individuals, as shown in Fig 1C. Table 1 contains informations of the identified proteins, including their accession number, matched peptides as well as theoretical and experimental molecular weight.

Bottom Line: Transthyretin amyloidosis is a conformational pathology characterized by the extracellular formation of amyloid deposits and the progressive impairment of the peripheral nervous system.Point mutations in this tetrameric plasma protein decrease its stability and are linked to disease onset and progression.We discovered, using a differential proteomics approach, that extracellular chaperones such as fibrinogen, clusterin, haptoglobin, alpha-1-anti-trypsin and 2-macroglobulin are overrepresented in transthyretin amyloidosis.

View Article: PubMed Central - PubMed

Affiliation: Centro de Química e Bioquímica, FCUL, Campo Grande, Lisboa, Portugal.

ABSTRACT
Transthyretin amyloidosis is a conformational pathology characterized by the extracellular formation of amyloid deposits and the progressive impairment of the peripheral nervous system. Point mutations in this tetrameric plasma protein decrease its stability and are linked to disease onset and progression. Since non-mutated transthyretin also forms amyloid in systemic senile amyloidosis and some mutation bearers are asymptomatic throughout their lives, non-genetic factors must also be involved in transthyretin amyloidosis. We discovered, using a differential proteomics approach, that extracellular chaperones such as fibrinogen, clusterin, haptoglobin, alpha-1-anti-trypsin and 2-macroglobulin are overrepresented in transthyretin amyloidosis. Our data shows that a complex network of extracellular chaperones are over represented in human plasma and we speculate that they act synergistically to cope with amyloid prone proteins. Proteostasis may thus be as important as point mutations in transthyretin amyloidosis.

No MeSH data available.


Related in: MedlinePlus