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http://dx.doi.org/10.5352/JLS.2011.21.12.1778

Modulation of G-CSF Secretion by Mutations of Non Alpha-Helical Region in N-Terminus  

Park, Jeong-Hae (Department of Clinical Laboratory Science, Dong Eui University)
Park, Jung-Ae (Department of Clinical Laboratory Science, Dong Eui University)
Kang, Seok-Woo (Department of Agricultural Biology, National Academy of Agricultural Science, RDA)
Goo, Tae-Won (Department of Agricultural Biology, National Academy of Agricultural Science, RDA)
Chung, Kyung-Tae (Department of Clinical Laboratory Science, Dong Eui University)
Publication Information
Journal of Life Science / v.21, no.12, 2011 , pp. 1778-1783 More about this Journal
Abstract
Hematopoietic cytokines regulate production of blood cells by stimulating proliferation and differentiation of bone marrow cells. Among these hematopoietic cytokines, called hematopoitic growth factors, glranulocyte-colony stimulating Factor (G-CSF), which regulates growth of neutrophils, is one of important therapeutic factors because cancer patients suffer with neutropenia which is severe reduction of neutrophils after chemotherapy. Two groups of recombinant G-CSF have approved and used for therapeutic purposes and many researches are still on-going to produce recombinant G-CSF by different techniques. We engineered human G-CSF with Bombyx specific endoplasmic reticulum (ER) signal sequence, therefore, secretion of human G-CSF protein was improved in Bombyx mori-origined cell line, Bm5. The Bombyx ER signal sequence and human G-CSF matured protein region chimera was further remodeled at the N-terminus of matured G-CSF protein to understand roles of N-terminus on outer cellular secretion and/or production. Three different mutants were generated deleting three amino acids in non alpha-helical region in N-terminus in order to scan important amino acids for G-CSF secretion. One of 3 different N-terminal deletion mutants showed dramatically reduction of secreted amount of G-CSF indicating its important role on secretion. The data suggest that remodeling in non alpha-helical region of N-terminus is also important for recombinant G-CSF production.
Keywords
G-CSF; Bombyx cell line; hematopoitic protein; deletion mutation;
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