Int. J. Mol. Sci. 2010, 11(10), 4051-4062; doi:10.3390/ijms11104051
Article

Carbonic Anhydrase I Is Recognized by an SOD1 Antibody upon Biotinylation of Human Spinal Cord Extracts

1 Department of Neuroscience, California Pacific Medical Center Research Institute, 475 Brannan Street, San Francisco, CA 94107, USA 2 Department of Cancer Research, California Pacific Medical Center Research Institute, 475 Brannan Street, San Francisco, CA 94107, USA 3 Department of Bioconformatics, California Pacific Medical Center Research Institute, 475 Brannan Street, San Francisco, CA 94107, USA 4 Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA Current Address: Department of Biological Sciences, Xi'an Jiaotong-Liverpool University, 111 Ren'ai Road, Suzhou Industrial Park, Suzhou 215123, Jiangsu Province, China.
* Authors to whom correspondence should be addressed.
Received: 16 September 2010; in revised form: 14 October 2010 / Accepted: 18 October 2010 / Published: 20 October 2010
(This article belongs to the Special Issue Biomarkers)
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Abstract: 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.
Keywords: mass spectrometry; proteomics; biotinylation; SOD1; ALS; carbonic anhydrase I

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MDPI and ACS Style

Liu, J.; Akhavan, A.; Lu, M.; Gruzman, A.; Lingappa, V.R.; An, J.; Bowser, R. Carbonic Anhydrase I Is Recognized by an SOD1 Antibody upon Biotinylation of Human Spinal Cord Extracts. Int. J. Mol. Sci. 2010, 11, 4051-4062.

AMA Style

Liu J, Akhavan A, Lu M, Gruzman A, Lingappa VR, An J, Bowser R. Carbonic Anhydrase I Is Recognized by an SOD1 Antibody upon Biotinylation of Human Spinal Cord Extracts. International Journal of Molecular Sciences. 2010; 11(10):4051-4062.

Chicago/Turabian Style

Liu, Jian; Akhavan, Armin; Lu, Mengde; Gruzman, Arie; Lingappa, Vishwanath R.; An, Jiyan; Bowser, Robert. 2010. "Carbonic Anhydrase I Is Recognized by an SOD1 Antibody upon Biotinylation of Human Spinal Cord Extracts." Int. J. Mol. Sci. 11, no. 10: 4051-4062.

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