Characterization of an ankyrin repeat-containing Shank2 isoform (Shank2E) in liver epithelial cells
Open accessAbstract
Shank proteins are a family of multidomain scaffolding proteins best known for their role in organizing the postsynaptic density region in neurons. Unlike Shank1 and Shank3, Shank2 [also known as Pro-SAP1 (proline-rich synapse-associated protein 1), CortBP1 (cortactin binding protein 1) or Spank-3] has been described as a truncated family member without an N-terminal ankyrin repeat domain. The present study utilized bioinformatics to demonstrate the presence of exons encoding ankyrin repeats in the region preceding the previously described Shank2 gene. cDNA sequencing of mRNA from epithelial cells revealed a novel spliceoform of Shank2, termed Shank2E, that encodes a predicted 200 kDa protein with six N-terminal ankyrin repeats. Shank2 mRNA from epithelial tissues was larger than transcripts in brain. Likewise, the apparent mass of Shank2 protein was larger in epithelial tissues (230 kDa) when compared with brain (165/180 kDa). Immunofluorescence and membrane fractionation found Shank2E concentrated at the apical membrane of liver epithelial cells. In cultured cholangiocytes, co-immunoprecipitation and detergent solubility studies revealed Shank2E complexed with actin and co-distributed with actin in detergent-insoluble lipid rafts. These findings indicate epithelial cells express an ankyrin repeat-containing Shank2 isoform, termed Shank2E, that is poised to co-ordinate actin-dependent events at the apical membrane.
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Differences in SHANK2 exon numbering across organisms in other reports likely reflect uncharacterized exons or alternative splicing.
“The difference in exon numbering for SHANK2 in different organisms in other reports is likely due to the pattern of uncharacterized exons or alternative splicing (Leblond et al., 2012; Lim et al., 1999; McWilliams et al., 2004).”
From Modeling Autism by SHANK Gene Mutations in Mice (2013) · cites this paper as McWilliams et al., 2004
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- Authors
- McWILLIAMS, R. R., GIDEY, E., FOUASSIER, L., WEED, S. A., & DOCTOR, R. B.
- Affiliations
- University of Colorado Health (US)
- Inserm (FR)
- Corresponding author
- GIDEY, Elizabeth
- Venue
- Biochemical Journal
- Publisher
- Portland Press Ltd.
- Published
- 2004-05-15
- Volume
- 380
- Issue
- 1
- Pages
- 181-191
- ISSN
- 0264-6021 (ISSN-L)
- 1470-8728
- PMC
- PMC1224161
- OpenAlex
- W2050182550
- Cited by
- 63
- References
- 42
- Citations by year
- 2026: 1 · 2024: 14 · 2023: 2 · 2022: 1 · 2020: 4 · 2018: 2 · 2017: 4 · 2015: 1 · 2014: 1 · 2013: 3 · 2012: 7
- FWCI
- 1.64
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Biochemical Journal (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01dk057729-05)
- National Institute of Diabetes and Digestive and Kidney Diseases (DK 57729)
- National Institute of Dental and Craniofacial Research (DE014364, K22DE014364, R01DE014578, DE014578)
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Keywords
- Shank2, ankyrin repeats, alternative splicing, liver epithelial cells, cholangiocytes, apical membrane, actin, lipid rafts, scaffold proteins, protein isoforms
- MeSH terms
- Amino Acid Sequence, Animals, Base Sequence, Bile Ducts, Intrahepatic, Carrier Proteins, Cells, Cultured, Epithelial Cells, Exons, Liver, Male, Actin Cytoskeleton, Molecular Sequence Data, Molecular Weight, Nerve Tissue Proteins, Organ Specificity, RNA, Messenger, Sequence Alignment, Cell Polarity, Ankyrin Repeat, Rats, Sprague-Dawley, Sequence Homology, Alternative Splicing, Protein Structure, Tertiary, Protein Isoforms, Membrane Microdomains, Hepatocytes, Rats
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