Carbonic Anhydrase I Is Recognized by an SOD1 Antibody upon Biotinylation of Human Spinal Cord Extracts
We recently reported the presence of a novel 32 kDa protein immunoreactive to a copper, zinc superoxide dismutase (SOD1) antibody within the spinal cord of patients with amyotrophic lateral sclerosis (ALS). This unique protein species was generated by biotinylation of spinal cord tissue extracts to...
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doaj-6bf963fa5a404ea98aca67d3c50af6012020-11-25T01:56:13ZengMDPI AGInternational Journal of Molecular Sciences1422-00672010-10-0111104051406210.3390/ijms11104051Carbonic Anhydrase I Is Recognized by an SOD1 Antibody upon Biotinylation of Human Spinal Cord ExtractsRobert BowserJiyan AnVishwanath R. LingappaMengde LuArmin AkhavanJian LiuArie GruzmanWe recently reported the presence of a novel 32 kDa protein immunoreactive to a copper, zinc superoxide dismutase (SOD1) antibody within the spinal cord of patients with amyotrophic lateral sclerosis (ALS). This unique protein species was generated by biotinylation of spinal cord tissue extracts to detect conformational changes of SOD1 specific to ALS patients. To further characterize this protein, we enriched the protein by column chromatography and determined its protein identity by mass spectrometry. The protein that gave rise to the 32 kDa species upon biotinylation was identified as carbonic anhydrase I (CA I). Biotinylation of CA I from ALS spinal cord resulted in the generation of a novel epitope recognized by the SOD1 antibody. This epitope could also be generated by biotinylation of extracts from cultured cells expressing human CA I. Peptide competition assays identified the amino acid sequence in carbonic anhydrase I responsible for binding the SOD1 antibody. We conclude that chemical modifications used to identify pathogenic protein conformations can lead to the identification of unanticipated proteins that may participate in disease pathogenesis. http://www.mdpi.com/1422-0067/11/10/4051/mass spectrometryproteomicsbiotinylationSOD1ALScarbonic anhydrase I |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Robert Bowser Jiyan An Vishwanath R. Lingappa Mengde Lu Armin Akhavan Jian Liu Arie Gruzman |
spellingShingle |
Robert Bowser Jiyan An Vishwanath R. Lingappa Mengde Lu Armin Akhavan Jian Liu Arie Gruzman Carbonic Anhydrase I Is Recognized by an SOD1 Antibody upon Biotinylation of Human Spinal Cord Extracts International Journal of Molecular Sciences mass spectrometry proteomics biotinylation SOD1 ALS carbonic anhydrase I |
author_facet |
Robert Bowser Jiyan An Vishwanath R. Lingappa Mengde Lu Armin Akhavan Jian Liu Arie Gruzman |
author_sort |
Robert Bowser |
title |
Carbonic Anhydrase I Is Recognized by an SOD1 Antibody upon Biotinylation of Human Spinal Cord Extracts |
title_short |
Carbonic Anhydrase I Is Recognized by an SOD1 Antibody upon Biotinylation of Human Spinal Cord Extracts |
title_full |
Carbonic Anhydrase I Is Recognized by an SOD1 Antibody upon Biotinylation of Human Spinal Cord Extracts |
title_fullStr |
Carbonic Anhydrase I Is Recognized by an SOD1 Antibody upon Biotinylation of Human Spinal Cord Extracts |
title_full_unstemmed |
Carbonic Anhydrase I Is Recognized by an SOD1 Antibody upon Biotinylation of Human Spinal Cord Extracts |
title_sort |
carbonic anhydrase i is recognized by an sod1 antibody upon biotinylation of human spinal cord extracts |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2010-10-01 |
description |
We recently reported the presence of a novel 32 kDa protein immunoreactive to a copper, zinc superoxide dismutase (SOD1) antibody within the spinal cord of patients with amyotrophic lateral sclerosis (ALS). This unique protein species was generated by biotinylation of spinal cord tissue extracts to detect conformational changes of SOD1 specific to ALS patients. To further characterize this protein, we enriched the protein by column chromatography and determined its protein identity by mass spectrometry. The protein that gave rise to the 32 kDa species upon biotinylation was identified as carbonic anhydrase I (CA I). Biotinylation of CA I from ALS spinal cord resulted in the generation of a novel epitope recognized by the SOD1 antibody. This epitope could also be generated by biotinylation of extracts from cultured cells expressing human CA I. Peptide competition assays identified the amino acid sequence in carbonic anhydrase I responsible for binding the SOD1 antibody. We conclude that chemical modifications used to identify pathogenic protein conformations can lead to the identification of unanticipated proteins that may participate in disease pathogenesis. |
topic |
mass spectrometry proteomics biotinylation SOD1 ALS carbonic anhydrase I |
url |
http://www.mdpi.com/1422-0067/11/10/4051/ |
work_keys_str_mv |
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