title: Full-length recombinant human SCF1-165 is more thermostable than the truncated SCF1-141 form.
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.
authors: 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.
authors: Yui-Ping
authors: YP
authors: Weng
authors: 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.
authors: Wen-Yen
authors: WY
authors: Ku
authors: Graduate Institute of Biological Science and Technology, Chung Hwa University of Medical Technology, Tainan, Taiwan.
authors: Ming-Han
authors: MH
authors: Wu
authors: Graduate Institute of Biological Science and Technology, Chung Hwa University of Medical Technology, Tainan, Taiwan.
authors: Ya-Li
authors: YL
authors: Tsai
authors: Graduate Institute of Biological Science and Technology, Chung Hwa University of Medical Technology, Tainan, Taiwan.
authors: Chi-Yu
authors: CY
authors: Chen
authors: Department of Biological Science and Technology, Chung Hwa University of Medical Technology, Tainan, Taiwan.
authors: Chun-An
authors: CA
authors: Kuo
authors: Institute of Biotechnology, Cheng Kung University, Tainan, Taiwan.
authors: Lynn L H
authors: LL
authors: Huang
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics; D002942:Circular Dichroism; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D011485:Protein Binding; D011487:Protein Conformation; D020033:Protein Isoforms / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D055550:Protein Stability; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D019089:Stem Cell Factor / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D013696:Temperature
chemical_list: D020033:Protein Isoforms; D011994:Recombinant Proteins; D019089:Stem Cell Factor