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Identification of cellular changes associated with increased production of human growth hormone in a recombinant Chinese hamster ovary cell line
Author(s) -
Van Dyk Derek D.,
Misztal David R.,
Wilkins Marc R.,
Mackintosh James A.,
Poljak Anne,
Varnai Jodie C.,
Teber Erdahl,
Walsh Bradley J.,
Gray Peter P.
Publication year - 2003
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200390023
Subject(s) - chinese hamster ovary cell , recombinant dna , proteomics , cell culture , biology , biochemistry , heat shock protein , hsp90 , microbiology and biotechnology , chemistry , receptor , genetics , gene
Abstract A proteomics approach was used to identify the proteins potentially implicated in the cellular response concomitant with elevated production levels of human growth hormone in a recombinant Chinese hamster ovary (CHO) cell line following exposure to 0.5 m M butyrate and 80 μ M zinc sulphate in the production media. This involved incorporation of two‐dimensional (2‐D) gel electrophoresis and protein identification by a combination of N ‐terminal sequencing, matrix‐assisted laser desorption/ionisation‐time of flight mass spectrometry, amino acid analysis and cross species database matching. From these identifications a CHO 2‐D reference map and annotated database have been established. Metabolic labelling and subsequent autoradiography showed the induction of a number of cellular proteins in response to the media additives butyrate and zinc sulphate. These were identified as GRP75, enolase and thioredoxin. The chaperone proteins GRP78, HSP90, GRP94 and HSP70 were not up‐regulated under these conditions.