Identification of the beta-dystroglycan binding epitope within the C-terminal region of alpha-dystroglycan
(2001) In European Journal of Biochemistry 268(16). p.4590-4597- Abstract
- Dystroglycan is a receptor for extracellular matrix proteins that plays a crucial role during embryogenesis in addition to adult tissue stabilization. A precursor product of a single gene is post-translationally cleaved to form two different subunits, alpha and beta. The extracellular alpha-dystroglycan is a membrane-associated, highly glycosylated protein that binds to various extracellular matrix molecules, whereas the transmembrane beta-dystroglycan binds, via its cytosolic domain, to dystrophin and many other proteins. alpha- and beta-Dystroglycan interact tightly but noncovalently. We have previously shown that the N-terminal region of beta-dystroglycan, beta-DG(654-750), binds to the C-terminal region of murine alpha-dystroglycan... (More)
- Dystroglycan is a receptor for extracellular matrix proteins that plays a crucial role during embryogenesis in addition to adult tissue stabilization. A precursor product of a single gene is post-translationally cleaved to form two different subunits, alpha and beta. The extracellular alpha-dystroglycan is a membrane-associated, highly glycosylated protein that binds to various extracellular matrix molecules, whereas the transmembrane beta-dystroglycan binds, via its cytosolic domain, to dystrophin and many other proteins. alpha- and beta-Dystroglycan interact tightly but noncovalently. We have previously shown that the N-terminal region of beta-dystroglycan, beta-DG(654-750), binds to the C-terminal region of murine alpha-dystroglycan independently from glycosylation. Preparing a series of deleted recombinant fragments and using solid-phase binding assays, the C-terminal sequence of alpha-dystroglycan containing the binding epitope for beta-dystroglycan has been defined more precisely. We found that a region of 36 amino acids, from position 550-585, is required for binding the extracellular region, amino acids 654-750 of beta-dystroglycan. Recently, a dystroglycan-like gene was identified in Drosophila that showed a moderate degree of conservation with vertebrate dystroglycan (31% identity, 48% similarity). Surprisingly, the Drosophila sequence contains a region showing a higher degree of identity and conservation (45% and 66%) that coincides with the 550-585 sequence of vertebrate alpha-dystroglycan. We have expressed this Drosophila dystroglycan fragment and measured its binding to the extracellular region of vertebrate (murine) beta-dystroglycan (Kd = 6 +/- 1 microM). These data confirm the proper identification of the beta-dystroglycan binding epitope and stress the importance of this region during evolution. This finding might help the rational design of dystroglycan-specific binding drugs, that could have important biomedical applications. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1121962
- author
- Sciandra, Francesca ; Schneider, Martina LU ; Giardina, Bruno ; Baumgartner, Stefan LU ; Petrucci, Tamara C. and Brancaccio, Andrea
- organization
- publishing date
- 2001
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- recombinant proteins, deletion mapping, protein–protein interaction, solid-phase binding assay, sequence alignment
- in
- European Journal of Biochemistry
- volume
- 268
- issue
- 16
- pages
- 4590 - 4597
- publisher
- Wiley-Blackwell
- external identifiers
-
- pmid:11502221
- scopus:0034832747
- ISSN
- 0014-2956
- DOI
- 10.1046/j.1432-1327.2001.02386.x
- language
- English
- LU publication?
- yes
- id
- ace2ecdb-2a1d-46a0-81cb-d81f998b17c8 (old id 1121962)
- date added to LUP
- 2016-04-01 16:08:56
- date last changed
- 2022-01-28 17:39:06
@article{ace2ecdb-2a1d-46a0-81cb-d81f998b17c8, abstract = {{Dystroglycan is a receptor for extracellular matrix proteins that plays a crucial role during embryogenesis in addition to adult tissue stabilization. A precursor product of a single gene is post-translationally cleaved to form two different subunits, alpha and beta. The extracellular alpha-dystroglycan is a membrane-associated, highly glycosylated protein that binds to various extracellular matrix molecules, whereas the transmembrane beta-dystroglycan binds, via its cytosolic domain, to dystrophin and many other proteins. alpha- and beta-Dystroglycan interact tightly but noncovalently. We have previously shown that the N-terminal region of beta-dystroglycan, beta-DG(654-750), binds to the C-terminal region of murine alpha-dystroglycan independently from glycosylation. Preparing a series of deleted recombinant fragments and using solid-phase binding assays, the C-terminal sequence of alpha-dystroglycan containing the binding epitope for beta-dystroglycan has been defined more precisely. We found that a region of 36 amino acids, from position 550-585, is required for binding the extracellular region, amino acids 654-750 of beta-dystroglycan. Recently, a dystroglycan-like gene was identified in Drosophila that showed a moderate degree of conservation with vertebrate dystroglycan (31% identity, 48% similarity). Surprisingly, the Drosophila sequence contains a region showing a higher degree of identity and conservation (45% and 66%) that coincides with the 550-585 sequence of vertebrate alpha-dystroglycan. We have expressed this Drosophila dystroglycan fragment and measured its binding to the extracellular region of vertebrate (murine) beta-dystroglycan (Kd = 6 +/- 1 microM). These data confirm the proper identification of the beta-dystroglycan binding epitope and stress the importance of this region during evolution. This finding might help the rational design of dystroglycan-specific binding drugs, that could have important biomedical applications.}}, author = {{Sciandra, Francesca and Schneider, Martina and Giardina, Bruno and Baumgartner, Stefan and Petrucci, Tamara C. and Brancaccio, Andrea}}, issn = {{0014-2956}}, keywords = {{recombinant proteins; deletion mapping; protein–protein interaction; solid-phase binding assay; sequence alignment}}, language = {{eng}}, number = {{16}}, pages = {{4590--4597}}, publisher = {{Wiley-Blackwell}}, series = {{European Journal of Biochemistry}}, title = {{Identification of the beta-dystroglycan binding epitope within the C-terminal region of alpha-dystroglycan}}, url = {{http://dx.doi.org/10.1046/j.1432-1327.2001.02386.x}}, doi = {{10.1046/j.1432-1327.2001.02386.x}}, volume = {{268}}, year = {{2001}}, }