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Full-Length Recombinant Human SCF(1-165) Is More Thermostable than the Truncated SCF(1-141) Form

Human stem cell factor initiates a diverse array of cellular responses, including hematopoiesis, cell proliferation, differentiation, migration and survival. To explore the relationship between its structure and function, we produced recombinant soluble human stem cell factor(1–165) (wild type) and...

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Autores principales: Weng, Yui-Ping, Ku, Wen-Yen, Wu, Ming-Han, Tsai, Ya-li, Chen, Chi-Yu, Kuo, Chun-An, Huang, Lynn L. H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4111497/
https://www.ncbi.nlm.nih.gov/pubmed/25061857
http://dx.doi.org/10.1371/journal.pone.0103251
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author Weng, Yui-Ping
Ku, Wen-Yen
Wu, Ming-Han
Tsai, Ya-li
Chen, Chi-Yu
Kuo, Chun-An
Huang, Lynn L. H.
author_facet Weng, Yui-Ping
Ku, Wen-Yen
Wu, Ming-Han
Tsai, Ya-li
Chen, Chi-Yu
Kuo, Chun-An
Huang, Lynn L. H.
author_sort Weng, Yui-Ping
collection PubMed
description Human stem cell factor initiates a diverse array of cellular responses, including hematopoiesis, cell proliferation, differentiation, migration and survival. To explore the relationship between its structure and function, we produced recombinant soluble human stem cell factor(1–165) (wild type) and human stem cell factor(1–141) (C-terminal truncated) in a yeast expression system and compared their biological activities and thermal stabilities. The biological activity of the two proteins was measured as a function of TF-1 cell viability and effects on downstream signaling targets after incubation. We found that these proteins enhanced cell viability and downstream signaling to a similar extent, in a dose-dependent manner. The biological activity of recombinant human stem cell factor(1–165) was significantly greater than that of recombinant human stem cell factor(1–141) after heating the proteins (100 ng/mL) at 25–110°C for 10 minutes (P<0.05 for all temperatures). In addition, circular dichroism spectral analysis indicated that β-sheet structures were altered in recombinant human stem cell factor(1–141) but not recombinant human stem cell factor(1–165) after heating at 90°C for 15 or 30 min. Molecular modeling and limited proteolytic digestion were also used to compare the thermo stability between human stem cell factor(1–165) and human stem cell factor(1–141). Together, these data indicate that stem cell factor(1–165) is more thermostable than stem cell factor(1–141).
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spelling pubmed-41114972014-07-29 Full-Length Recombinant Human SCF(1-165) Is More Thermostable than the Truncated SCF(1-141) Form Weng, Yui-Ping Ku, Wen-Yen Wu, Ming-Han Tsai, Ya-li Chen, Chi-Yu Kuo, Chun-An Huang, Lynn L. H. PLoS One Research Article Human stem cell factor initiates a diverse array of cellular responses, including hematopoiesis, cell proliferation, differentiation, migration and survival. To explore the relationship between its structure and function, we produced recombinant soluble human stem cell factor(1–165) (wild type) and human stem cell factor(1–141) (C-terminal truncated) in a yeast expression system and compared their biological activities and thermal stabilities. The biological activity of the two proteins was measured as a function of TF-1 cell viability and effects on downstream signaling targets after incubation. We found that these proteins enhanced cell viability and downstream signaling to a similar extent, in a dose-dependent manner. The biological activity of recombinant human stem cell factor(1–165) was significantly greater than that of recombinant human stem cell factor(1–141) after heating the proteins (100 ng/mL) at 25–110°C for 10 minutes (P<0.05 for all temperatures). In addition, circular dichroism spectral analysis indicated that β-sheet structures were altered in recombinant human stem cell factor(1–141) but not recombinant human stem cell factor(1–165) after heating at 90°C for 15 or 30 min. Molecular modeling and limited proteolytic digestion were also used to compare the thermo stability between human stem cell factor(1–165) and human stem cell factor(1–141). Together, these data indicate that stem cell factor(1–165) is more thermostable than stem cell factor(1–141). Public Library of Science 2014-07-25 /pmc/articles/PMC4111497/ /pubmed/25061857 http://dx.doi.org/10.1371/journal.pone.0103251 Text en © 2014 Weng et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Weng, Yui-Ping
Ku, Wen-Yen
Wu, Ming-Han
Tsai, Ya-li
Chen, Chi-Yu
Kuo, Chun-An
Huang, Lynn L. H.
Full-Length Recombinant Human SCF(1-165) Is More Thermostable than the Truncated SCF(1-141) Form
title Full-Length Recombinant Human SCF(1-165) Is More Thermostable than the Truncated SCF(1-141) Form
title_full Full-Length Recombinant Human SCF(1-165) Is More Thermostable than the Truncated SCF(1-141) Form
title_fullStr Full-Length Recombinant Human SCF(1-165) Is More Thermostable than the Truncated SCF(1-141) Form
title_full_unstemmed Full-Length Recombinant Human SCF(1-165) Is More Thermostable than the Truncated SCF(1-141) Form
title_short Full-Length Recombinant Human SCF(1-165) Is More Thermostable than the Truncated SCF(1-141) Form
title_sort full-length recombinant human scf(1-165) is more thermostable than the truncated scf(1-141) form
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4111497/
https://www.ncbi.nlm.nih.gov/pubmed/25061857
http://dx.doi.org/10.1371/journal.pone.0103251
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