Open AccessFull-Length Recombinant Human SCF1-165 Is More Thermostable than the Truncated SCF1-141 Form
Author(s) -
Yu-I Weng,
Wen Yen Ku,
Ming Han Wu,
Ya Li Tsai,
Chi Yu Chen,
Chun An Kuo,
Lynn Ling-Huei Huang
Publication year - 2014
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0103251
Subject(s) - stem cell , recombinant dna , biology , microbiology and biotechnology , cell , viability assay , cell growth , biochemistry , gene
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 .