| RRC ID |
58054
|
| 著者 |
Sugawara Y, Hamada K, Yamada Y, Kumai J, Kanagawa M, Kobayashi K, Toda T, Negishi Y, Katagiri F, Hozumi K, Nomizu M, Kikkawa Y.
|
| タイトル |
Characterization of dystroglycan binding in adhesion of human induced pluripotent stem cells to laminin-511 E8 fragment.
|
| ジャーナル |
Sci Rep
|
| Abstract |
Human induced pluripotent stem cells (hiPSCs) grow indefinitely in culture and have the potential to regenerate various tissues. In the development of cell culture systems, a fragment of laminin-511 (LM511-E8) was found to improve the proliferation of stem cells. The adhesion of undifferentiated cells to LM511-E8 is mainly mediated through integrin α6β1. However, the involvement of non-integrin receptors remains unknown in stem cell culture using LM511-E8. Here, we show that dystroglycan (DG) is strongly expressed in hiPSCs. The fully glycosylated DG is functionally active for laminin binding, and although it has been suggested that LM511-E8 lacks DG binding sites, the fragment does weakly bind to DG. We further identified the DG binding sequence in LM511-E8, using synthetic peptides, of which, hE8A5-20 (human laminin α5 2688-2699: KTLPQLLAKLSI) derived from the laminin coiled-coil domain, exhibited DG binding affinity and cell adhesion activity. Deletion and mutation studies show that LLAKLSI is the active core sequence of hE8A5-20, and that, K2696 is a critical amino acid for DG binding. We further demonstrated that hiPSCs adhere to hE8A5-20-conjugated chitosan matrices. The amino acid sequence of DG binding peptides would be useful to design substrata for culture system of undifferentiated and differentiated stem cells.
|
| 巻・号 |
9(1)
|
| ページ |
13037
|
| 公開日 |
2019-9-10
|
| DOI |
10.1038/s41598-019-49669-x
|
| PII |
10.1038/s41598-019-49669-x
|
| PMID |
31506597
|
| PMC |
PMC6737067
|
| MeSH |
Amino Acid Sequence
Biomarkers
Cell Adhesion*
Cells, Cultured
Dystroglycans / metabolism*
Extracellular Matrix
Fluorescent Antibody Technique
Humans
Induced Pluripotent Stem Cells / metabolism*
Laminin / chemistry
Laminin / metabolism*
Peptide Fragments / chemistry
Peptide Fragments / metabolism*
Protein Binding
Protein Transport
|
| IF |
3.998
|
| 引用数 |
0
|
| リソース情報 |
| ヒト・動物細胞 |
201B7(HPS0063)
454E2(HPS0077) |
