Full-length recombinant human SCF1-165 is more thermostable than the truncated SCF1-141 form

Yui-Ping Weng; Wen-Yen Ku; Ming-Han Wu; Ya-Li Tsai; Chi-Yu Chen; Chun-An Kuo; Lynn L H Huang

doi:10.1371/journal.pone.0103251

Full-length recombinant human SCF1-165 is more thermostable than the truncated SCF1-141 form

PLoS One. 2014 Jul 25;9(7):e103251. doi: 10.1371/journal.pone.0103251. eCollection 2014.

Authors

Yui-Ping Weng¹, Wen-Yen Ku¹, Ming-Han Wu², Ya-Li Tsai², Chi-Yu Chen², Chun-An Kuo³, Lynn L H Huang⁴

Affiliations

¹ Graduate Institute of Biological Science and Technology, Chung Hwa University of Medical Technology, Tainan, Taiwan; Department of Biological Science and Technology, Chung Hwa University of Medical Technology, Tainan, Taiwan.
² Graduate Institute of Biological Science and Technology, Chung Hwa University of Medical Technology, Tainan, Taiwan.
³ Department of Biological Science and Technology, Chung Hwa University of Medical Technology, Tainan, Taiwan.
⁴ Institute of Biotechnology, Cheng Kung University, Tainan, Taiwan.

Abstract

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 factor1-165 (wild type) and human stem cell factor1-141 (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 factor1-165 was significantly greater than that of recombinant human stem cell factor1-141 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 factor1-141 but not recombinant human stem cell factor1-165 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 factor1-165 and human stem cell factor1-141. Together, these data indicate that stem cell factor1-165 is more thermostable than stem cell factor1-141.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line, Tumor
Cell Survival / genetics
Circular Dichroism
Humans
Kinetics
Models, Molecular
Molecular Dynamics Simulation
Protein Binding
Protein Conformation
Protein Isoforms / biosynthesis
Protein Isoforms / chemistry
Protein Isoforms / genetics*
Protein Stability*
Recombinant Proteins / biosynthesis*
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Stem Cell Factor / biosynthesis*
Stem Cell Factor / chemistry
Stem Cell Factor / genetics
Temperature

Substances

Protein Isoforms
Recombinant Proteins
Stem Cell Factor

Grants and funding

This work was partially supported by Research Grants (NSC 102-2325-B-006 -012) from the Ministry of Science and Technology. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.