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Ankyrin-binding domain of CD44(GP85) is required for the expression of hyaluronic acid-mediated adhesion function
GP85 is one of the most common hemopoietic isoforms of the cell adhesion molecule, CD44. CD44(GP85) is known to contain at least one ankyrin-binding site within its 70 aa cytoplasmic domain and to bind hyaluronic acid (HA) with its extracellular domain. In this study we have mapped the ankyrin-bindi...
Formato: | Texto |
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Lenguaje: | English |
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The Rockefeller University Press
1994
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2120123/ https://www.ncbi.nlm.nih.gov/pubmed/7519619 |
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collection | PubMed |
description | GP85 is one of the most common hemopoietic isoforms of the cell adhesion molecule, CD44. CD44(GP85) is known to contain at least one ankyrin-binding site within its 70 aa cytoplasmic domain and to bind hyaluronic acid (HA) with its extracellular domain. In this study we have mapped the ankyrin-binding domain of CD44(GP85) by deleting various portions of the cytoplasmic region followed by expression of these truncated cDNAs in COS cells. The results of these experiments indicate that the ankyrin-binding domain resides between amino acids 305 and 355. Biochemical analyses, using competition binding assays and a synthetic peptide (NGGNGT-VEDRKPSEL) containing 15 aa between aa 305 and aa 320, support the conclusion that this region is required for ankryin binding. Furthermore, we have constructed a fusion protein in which this 15 aa sequence of CD44(GP85) is transplanted onto another transmembrane protein which does not bind ankyrin. Our results show that this fusion protein acquires the ability to bind ankyrin confirming that the sequence (306NGGNGTVEDRKPSE320L) is a critical part of the ankryin-binding domain of CD44(GP85). In addition, we have demonstrated that deletion of this 15 aa ankyrin-binding sequence from CD44(GP85) results in a drastic reduction (> or = 90%) of HA-binding and HA-mediated cell adhesion. These findings strongly suggest that ankyrin binding to the cytoplasmic domain of CD44(GP85) plays a pivotal role in regulating hyaluronic acid-mediated cell-cell and cell- extracellular matrix interactions. |
format | Text |
id | pubmed-2120123 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1994 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-21201232008-05-01 Ankyrin-binding domain of CD44(GP85) is required for the expression of hyaluronic acid-mediated adhesion function J Cell Biol Articles GP85 is one of the most common hemopoietic isoforms of the cell adhesion molecule, CD44. CD44(GP85) is known to contain at least one ankyrin-binding site within its 70 aa cytoplasmic domain and to bind hyaluronic acid (HA) with its extracellular domain. In this study we have mapped the ankyrin-binding domain of CD44(GP85) by deleting various portions of the cytoplasmic region followed by expression of these truncated cDNAs in COS cells. The results of these experiments indicate that the ankyrin-binding domain resides between amino acids 305 and 355. Biochemical analyses, using competition binding assays and a synthetic peptide (NGGNGT-VEDRKPSEL) containing 15 aa between aa 305 and aa 320, support the conclusion that this region is required for ankryin binding. Furthermore, we have constructed a fusion protein in which this 15 aa sequence of CD44(GP85) is transplanted onto another transmembrane protein which does not bind ankyrin. Our results show that this fusion protein acquires the ability to bind ankyrin confirming that the sequence (306NGGNGTVEDRKPSE320L) is a critical part of the ankryin-binding domain of CD44(GP85). In addition, we have demonstrated that deletion of this 15 aa ankyrin-binding sequence from CD44(GP85) results in a drastic reduction (> or = 90%) of HA-binding and HA-mediated cell adhesion. These findings strongly suggest that ankyrin binding to the cytoplasmic domain of CD44(GP85) plays a pivotal role in regulating hyaluronic acid-mediated cell-cell and cell- extracellular matrix interactions. The Rockefeller University Press 1994-08-02 /pmc/articles/PMC2120123/ /pubmed/7519619 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Articles Ankyrin-binding domain of CD44(GP85) is required for the expression of hyaluronic acid-mediated adhesion function |
title | Ankyrin-binding domain of CD44(GP85) is required for the expression of hyaluronic acid-mediated adhesion function |
title_full | Ankyrin-binding domain of CD44(GP85) is required for the expression of hyaluronic acid-mediated adhesion function |
title_fullStr | Ankyrin-binding domain of CD44(GP85) is required for the expression of hyaluronic acid-mediated adhesion function |
title_full_unstemmed | Ankyrin-binding domain of CD44(GP85) is required for the expression of hyaluronic acid-mediated adhesion function |
title_short | Ankyrin-binding domain of CD44(GP85) is required for the expression of hyaluronic acid-mediated adhesion function |
title_sort | ankyrin-binding domain of cd44(gp85) is required for the expression of hyaluronic acid-mediated adhesion function |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2120123/ https://www.ncbi.nlm.nih.gov/pubmed/7519619 |