Identification of cellular changes associated with increased production of human growth hormone in a recombinant Chinese hamster ovary cell line

Derek D. Van Dyk*, David R. Misztal, Marc R. Wilkins, James A. Mackintosh, Anne Poljak, Jodie C. Varnai, Erdahl Teber, Bradley J. Walsh, Peter P. Gray

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

46 Citations (Scopus)

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 mM butyrate and 80 μM 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.

Original languageEnglish
Pages (from-to)147-156
Number of pages10
JournalProteomics
Volume3
Issue number2
DOIs
Publication statusPublished - 1 Feb 2003

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