Ribosylation rapidly induces alpha-synuclein to form highly cytotoxic molten globules of advanced glycation end products.
BACKGROUND: Alpha synuclein (alpha-Syn) is the main component of Lewy bodies which are associated with several neurodegenerative diseases such as Parkinson's disease. While the glycation with D-glucose that results in alpha-Syn misfold and aggregation has been studied, the effects of glycation...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2010-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC2816216?pdf=render |
| _version_ | 1828766504247623680 |
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| author | Lan Chen Yan Wei Xueqing Wang Rongqiao He |
| author_facet | Lan Chen Yan Wei Xueqing Wang Rongqiao He |
| author_sort | Lan Chen |
| collection | DOAJ |
| description | BACKGROUND: Alpha synuclein (alpha-Syn) is the main component of Lewy bodies which are associated with several neurodegenerative diseases such as Parkinson's disease. While the glycation with D-glucose that results in alpha-Syn misfold and aggregation has been studied, the effects of glycation with D-ribose on alpha-Syn have not been investigated. METHODOLOGY/PRINCIPAL FINDINGS: Here, we show that ribosylation induces alpha-Syn misfolding and generates advanced glycation end products (AGEs) which form protein molten globules with high cytotoxcity. Results from native- and SDS-PAGE showed that D-ribose reacted rapidly with alpha-Syn, leading to dimerization and polymerization. Trypsin digestion and sequencing analysis revealed that during ribosylation the lysinyl residues (K(58), K(60), K(80), K(96), K(97) and K(102)) in the C-terminal region reacted more quickly with D-ribose than those of the N-terminal region. Using Western blotting, AGEs resulting from the glycation of alpha-Syn were observed within 24 h in the presence of D-ribose, but were not observed in the presence of D-glucose. Changes in fluorescence at 410 nm demonstrated again that AGEs were formed during early ribosylation. Changes in the secondary structure of ribosylated alpha-Syn were not clearly detected by CD spectrometry in studies on protein conformation. However, intrinsic fluorescence at 310 nm decreased markedly in the presence of D-ribose. Observations with atomic force microscopy showed that the surface morphology of glycated alpha-Syn looked like globular aggregates. thioflavin T (ThT) fluorescence increased during alpha-Syn incubation regardless of ribosylation. As incubation time increased, ribosylation of alpha-Syn resulted in a blue-shift (approximately 100 nm) in the fluorescence of ANS. The light scattering intensity of ribosylated alpha-Syn was not markedly different from native alpha-Syn, suggesting that ribosylated alpha-Syn is present as molten protein globules. Ribosylated products had a high cytotoxicity to SH-SY5Y cells, leading to LDH release and increase in the levels of reactive oxygen species (ROS). CONCLUSIONS/SIGNIFICANCE: alpha-Syn is rapidly glycated in the presence of D-ribose generating molten globule-like aggregations which cause cell oxidative stress and result in high cytotoxicity. |
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| format | Article |
| id | doaj.art-2a40cd6b19d34ac0b7f345375c20cb15 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-11T07:07:26Z |
| publishDate | 2010-01-01 |
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| spelling | doaj.art-2a40cd6b19d34ac0b7f345375c20cb152022-12-22T01:16:26ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-01-0152e905210.1371/journal.pone.0009052Ribosylation rapidly induces alpha-synuclein to form highly cytotoxic molten globules of advanced glycation end products.Lan ChenYan WeiXueqing WangRongqiao HeBACKGROUND: Alpha synuclein (alpha-Syn) is the main component of Lewy bodies which are associated with several neurodegenerative diseases such as Parkinson's disease. While the glycation with D-glucose that results in alpha-Syn misfold and aggregation has been studied, the effects of glycation with D-ribose on alpha-Syn have not been investigated. METHODOLOGY/PRINCIPAL FINDINGS: Here, we show that ribosylation induces alpha-Syn misfolding and generates advanced glycation end products (AGEs) which form protein molten globules with high cytotoxcity. Results from native- and SDS-PAGE showed that D-ribose reacted rapidly with alpha-Syn, leading to dimerization and polymerization. Trypsin digestion and sequencing analysis revealed that during ribosylation the lysinyl residues (K(58), K(60), K(80), K(96), K(97) and K(102)) in the C-terminal region reacted more quickly with D-ribose than those of the N-terminal region. Using Western blotting, AGEs resulting from the glycation of alpha-Syn were observed within 24 h in the presence of D-ribose, but were not observed in the presence of D-glucose. Changes in fluorescence at 410 nm demonstrated again that AGEs were formed during early ribosylation. Changes in the secondary structure of ribosylated alpha-Syn were not clearly detected by CD spectrometry in studies on protein conformation. However, intrinsic fluorescence at 310 nm decreased markedly in the presence of D-ribose. Observations with atomic force microscopy showed that the surface morphology of glycated alpha-Syn looked like globular aggregates. thioflavin T (ThT) fluorescence increased during alpha-Syn incubation regardless of ribosylation. As incubation time increased, ribosylation of alpha-Syn resulted in a blue-shift (approximately 100 nm) in the fluorescence of ANS. The light scattering intensity of ribosylated alpha-Syn was not markedly different from native alpha-Syn, suggesting that ribosylated alpha-Syn is present as molten protein globules. Ribosylated products had a high cytotoxicity to SH-SY5Y cells, leading to LDH release and increase in the levels of reactive oxygen species (ROS). CONCLUSIONS/SIGNIFICANCE: alpha-Syn is rapidly glycated in the presence of D-ribose generating molten globule-like aggregations which cause cell oxidative stress and result in high cytotoxicity.http://europepmc.org/articles/PMC2816216?pdf=render |
| spellingShingle | Lan Chen Yan Wei Xueqing Wang Rongqiao He Ribosylation rapidly induces alpha-synuclein to form highly cytotoxic molten globules of advanced glycation end products. PLoS ONE |
| title | Ribosylation rapidly induces alpha-synuclein to form highly cytotoxic molten globules of advanced glycation end products. |
| title_full | Ribosylation rapidly induces alpha-synuclein to form highly cytotoxic molten globules of advanced glycation end products. |
| title_fullStr | Ribosylation rapidly induces alpha-synuclein to form highly cytotoxic molten globules of advanced glycation end products. |
| title_full_unstemmed | Ribosylation rapidly induces alpha-synuclein to form highly cytotoxic molten globules of advanced glycation end products. |
| title_short | Ribosylation rapidly induces alpha-synuclein to form highly cytotoxic molten globules of advanced glycation end products. |
| title_sort | ribosylation rapidly induces alpha synuclein to form highly cytotoxic molten globules of advanced glycation end products |
| url | http://europepmc.org/articles/PMC2816216?pdf=render |
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