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Title: Copper and Zinc Metallation Status of Copper Zinc Superoxide Dismutase form Amyotrophic Lateral Sclerosis Transgenic Mice

Abstract

Mutations in the metalloenzyme copper-zinc superoxide dismutase (SOD1) cause one form of familial amyotrophic lateral sclerosis (ALS), and metals are suspected to play a pivotal role in ALS pathology. To learn more about metals in ALS, we determined the metallation states of human wild-type or mutant (G37R, G93A, and H46R/H48Q) SOD1 proteins from SOD1-ALS transgenic mice spinal cords. SOD1 was gently extracted from spinal cord and separated into insoluble (aggregated) and soluble (supernatant) fractions, and then metallation states were determined by HPLC inductively coupled plasma MS. Insoluble SOD1-rich fractions were not enriched in copper and zinc. However, the soluble mutant and WT SOD1s were highly metallated except for the metal-binding-region mutant H46R/H48Q, which did not bind any copper. Due to the stability conferred by high metallation of G37R and G93A, it is unlikely that these soluble SOD1s are prone to aggregation in vivo, supporting the hypothesis that immature nascent SOD1 is the substrate for aggregation. We also investigated the effect of SOD1 overexpression and disease on metal homeostasis in spinal cord cross-sections of SOD1-ALS mice using synchrotron-based x-ray fluorescence microscopy. In each mouse genotype, except for the H46R/H48Q mouse, we found a redistribution of copper between gray and white mattersmore » correlated to areas of high SOD1. Interestingly, a disease-specific increase of zinc was observed in the white matter for all mutant SOD1 mice. Together these data provide a picture of copper and zinc in the cell as well as highlight the importance of these metals in understanding SOD1-ALS pathology.« less

Authors:
; ; ; ; ; ; ;
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source
Sponsoring Org.:
DOE - OFFICE OF SCIENCE
OSTI Identifier:
1019462
Report Number(s):
BNL-94908-2011-JA
R&D Project: LS001; KC0401030; TRN: US201115%%136
DOE Contract Number:  
DE-AC02-98CH10886
Resource Type:
Journal Article
Journal Name:
The Journal of Biological Chemistry
Additional Journal Information:
Journal Volume: 286; Journal Issue: 4
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; COPPER; CROSS SECTIONS; DISEASES; FLUORESCENCE; GENOTYPE; HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY; HOMEOSTASIS; HYPOTHESIS; IN VIVO; MICE; MICROSCOPY; MUTANTS; MUTATIONS; PATHOLOGY; PROTEINS; SPINAL CORD; STABILITY; SUBSTRATES; SUPEROXIDE DISMUTASE; TRANSGENIC MICE; ZINC; national synchrotron light source

Citation Formats

Lelie, H L, Miller, L, Liba, A, Bourassa, M W, Chattopadhyay, M, Chan, P K, Gralla, E B, Borchelt, D R, and et al. Copper and Zinc Metallation Status of Copper Zinc Superoxide Dismutase form Amyotrophic Lateral Sclerosis Transgenic Mice. United States: N. p., 2010. Web.
Lelie, H L, Miller, L, Liba, A, Bourassa, M W, Chattopadhyay, M, Chan, P K, Gralla, E B, Borchelt, D R, & et al. Copper and Zinc Metallation Status of Copper Zinc Superoxide Dismutase form Amyotrophic Lateral Sclerosis Transgenic Mice. United States.
Lelie, H L, Miller, L, Liba, A, Bourassa, M W, Chattopadhyay, M, Chan, P K, Gralla, E B, Borchelt, D R, and et al. 2010. "Copper and Zinc Metallation Status of Copper Zinc Superoxide Dismutase form Amyotrophic Lateral Sclerosis Transgenic Mice". United States.
@article{osti_1019462,
title = {Copper and Zinc Metallation Status of Copper Zinc Superoxide Dismutase form Amyotrophic Lateral Sclerosis Transgenic Mice},
author = {Lelie, H L and Miller, L and Liba, A and Bourassa, M W and Chattopadhyay, M and Chan, P K and Gralla, E B and Borchelt, D R and et al},
abstractNote = {Mutations in the metalloenzyme copper-zinc superoxide dismutase (SOD1) cause one form of familial amyotrophic lateral sclerosis (ALS), and metals are suspected to play a pivotal role in ALS pathology. To learn more about metals in ALS, we determined the metallation states of human wild-type or mutant (G37R, G93A, and H46R/H48Q) SOD1 proteins from SOD1-ALS transgenic mice spinal cords. SOD1 was gently extracted from spinal cord and separated into insoluble (aggregated) and soluble (supernatant) fractions, and then metallation states were determined by HPLC inductively coupled plasma MS. Insoluble SOD1-rich fractions were not enriched in copper and zinc. However, the soluble mutant and WT SOD1s were highly metallated except for the metal-binding-region mutant H46R/H48Q, which did not bind any copper. Due to the stability conferred by high metallation of G37R and G93A, it is unlikely that these soluble SOD1s are prone to aggregation in vivo, supporting the hypothesis that immature nascent SOD1 is the substrate for aggregation. We also investigated the effect of SOD1 overexpression and disease on metal homeostasis in spinal cord cross-sections of SOD1-ALS mice using synchrotron-based x-ray fluorescence microscopy. In each mouse genotype, except for the H46R/H48Q mouse, we found a redistribution of copper between gray and white matters correlated to areas of high SOD1. Interestingly, a disease-specific increase of zinc was observed in the white matter for all mutant SOD1 mice. Together these data provide a picture of copper and zinc in the cell as well as highlight the importance of these metals in understanding SOD1-ALS pathology.},
doi = {},
url = {https://www.osti.gov/biblio/1019462}, journal = {The Journal of Biological Chemistry},
number = 4,
volume = 286,
place = {United States},
year = {Fri Sep 24 00:00:00 EDT 2010},
month = {Fri Sep 24 00:00:00 EDT 2010}
}