Identification of cellular changes associated with increased production of human growth hormone in a recombinant Chinese hamster ovary cell line.
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 microM 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.
Van Dyk
Derek D
DD
Australian Proteome Analysis Facility, Macquarie University, Sydney, Australia. dvandyk@proteome.org.au
Misztal
David R
DR
Wilkins
Marc R
MR
Mackintosh
James A
JA
Poljak
Anne
A
Varnai
Jodie C
JC
Teber
Erdahl
E
Walsh
Bradley J
BJ
Gray
Peter P
PP
D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D016466:CHO Cells; D006224:Cricetinae; D003470:Culture Media; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D019992:Databases as Topic; D015180:Electrophoresis, Gel, Two-Dimensional; D000091342:Endoplasmic Reticulum Chaperone BiP; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D019382:Human Growth Hormone / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D008565:Membrane Proteins / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D017434:Protein Structure, Tertiary; D020543:Proteome; D013045:Species Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013879:Thioredoxins / Q000378:metabolism; D015032:Zinc / Q000494:pharmacology
D000596:Amino Acids; D002087:Butyrates; D003470:Culture Media; D016895:Culture Media, Serum-Free; D000091342:Endoplasmic Reticulum Chaperone BiP; D018840:HSP70 Heat-Shock Proteins; C000717247:HSPA5 protein, human; D008565:Membrane Proteins; D020543:Proteome; C018551:glucose-regulated proteins; D019382:Human Growth Hormone; D013879:Thioredoxins; D010751:Phosphopyruvate Hydratase; D015032:Zinc