Mechanism of Action of Secreted Newt Anterior Gradient Protein.
Anterior gradient (AG) proteins have a thioredoxin fold and are targeted to the secretory pathway where they may act in the ER, as well as after secretion into the extracellular space. A newt member of the family (nAG) was previously identified as interacting with the GPI-anchored salamander-specifi...
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Public Library of Science (PLoS)
2016-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0154176&type=printable |
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| author | Kathrin S Grassme Acely Garza-Garcia Jean-Paul Delgado James W Godwin Anoop Kumar Phillip B Gates Paul C Driscoll Jeremy P Brockes |
| author_facet | Kathrin S Grassme Acely Garza-Garcia Jean-Paul Delgado James W Godwin Anoop Kumar Phillip B Gates Paul C Driscoll Jeremy P Brockes |
| author_sort | Kathrin S Grassme |
| collection | DOAJ |
| description | Anterior gradient (AG) proteins have a thioredoxin fold and are targeted to the secretory pathway where they may act in the ER, as well as after secretion into the extracellular space. A newt member of the family (nAG) was previously identified as interacting with the GPI-anchored salamander-specific three-finger protein called Prod1. Expression of nAG has been implicated in the nerve dependence of limb regeneration in salamanders, and nAG acted as a growth factor for cultured newt limb blastemal (progenitor) cells, but the mechanism of action was not understood. Here we show that addition of a peptide antibody to Prod1 specifically inhibit the proliferation of blastema cells, suggesting that Prod1 acts as a cell surface receptor for secreted nAG, leading to S phase entry. Mutation of the single cysteine residue in the canonical active site of nAG to alanine or serine leads to protein degradation, but addition of residues at the C terminus stabilises the secreted protein. The mutation of the cysteine residue led to no detectable activity on S phase entry in cultured newt limb blastemal cells. In addition, our phylogenetic analyses have identified a new Caudata AG protein called AG4. A comparison of the AG proteins in a cell culture assay indicates that nAG secretion is significantly higher than AGR2 or AG4, suggesting that this property may vary in different members of the family. |
| format | Article |
| id | doaj-art-77becb1e71eb488abd4def8377a9719c |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-77becb1e71eb488abd4def8377a9719c2025-08-20T03:26:10ZengPublic Library of Science (PLoS)PLoS ONE1932-62032016-01-01114e015417610.1371/journal.pone.0154176Mechanism of Action of Secreted Newt Anterior Gradient Protein.Kathrin S GrassmeAcely Garza-GarciaJean-Paul DelgadoJames W GodwinAnoop KumarPhillip B GatesPaul C DriscollJeremy P BrockesAnterior gradient (AG) proteins have a thioredoxin fold and are targeted to the secretory pathway where they may act in the ER, as well as after secretion into the extracellular space. A newt member of the family (nAG) was previously identified as interacting with the GPI-anchored salamander-specific three-finger protein called Prod1. Expression of nAG has been implicated in the nerve dependence of limb regeneration in salamanders, and nAG acted as a growth factor for cultured newt limb blastemal (progenitor) cells, but the mechanism of action was not understood. Here we show that addition of a peptide antibody to Prod1 specifically inhibit the proliferation of blastema cells, suggesting that Prod1 acts as a cell surface receptor for secreted nAG, leading to S phase entry. Mutation of the single cysteine residue in the canonical active site of nAG to alanine or serine leads to protein degradation, but addition of residues at the C terminus stabilises the secreted protein. The mutation of the cysteine residue led to no detectable activity on S phase entry in cultured newt limb blastemal cells. In addition, our phylogenetic analyses have identified a new Caudata AG protein called AG4. A comparison of the AG proteins in a cell culture assay indicates that nAG secretion is significantly higher than AGR2 or AG4, suggesting that this property may vary in different members of the family.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0154176&type=printable |
| spellingShingle | Kathrin S Grassme Acely Garza-Garcia Jean-Paul Delgado James W Godwin Anoop Kumar Phillip B Gates Paul C Driscoll Jeremy P Brockes Mechanism of Action of Secreted Newt Anterior Gradient Protein. PLoS ONE |
| title | Mechanism of Action of Secreted Newt Anterior Gradient Protein. |
| title_full | Mechanism of Action of Secreted Newt Anterior Gradient Protein. |
| title_fullStr | Mechanism of Action of Secreted Newt Anterior Gradient Protein. |
| title_full_unstemmed | Mechanism of Action of Secreted Newt Anterior Gradient Protein. |
| title_short | Mechanism of Action of Secreted Newt Anterior Gradient Protein. |
| title_sort | mechanism of action of secreted newt anterior gradient protein |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0154176&type=printable |
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