Characterization of growth hormone releasing factor analog expression in Saccharomyces cerevisiae.
An analog of growth hormone releasing factor (GRF), [Leu27]GRF(1-40)-OH, has been expressed and secreted in Saccharomyces cerevisiae under the control of the alpha-factor gene promoter and prepro sequence. A single pair of consecutive basic residues served as a processing site between the alpha-factor sequences and the GRF sequences. [Leu27]GRF(1-40)-OH from fermentor broth containing 20-30 mg/L of immunoreactive peptides was shown to be correctly processed and to possess biological activity as measured in vitro and in vivo. Additional peptides purified from broth appear to result from proteolytic degradation of the original translation product. Analysis of the amino acid compositions and sequences of these peptides suggests that processing enzymes may be responsible for some of the degradation.
Craig
W S
WS
Salk Institute Biotechnology/Industrial Associates, San Diego, CA.
Wondrack
L
L
Siegel
R
R
Patthi
S
S
Davis
G R
GR
Velicelebi
G
G
Mowles
T F
TF
Thill
G P
GP
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001681:Biological Assay; D002478:Cells, Cultured; D015870:Gene Expression; D005822:Genetic Vectors; D013007:Growth Hormone-Releasing Hormone / Q000031:analogs & derivatives* / Q000235:genetics; D000072235:Mating Factor; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics* / Q000378:metabolism; D010455:Peptides / Q000235:genetics; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D021382:Protein Sorting Signals / Q000235:genetics; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014162:Transfection / Q000235:genetics
D000596:Amino Acids; D010446:Peptide Fragments; D010455:Peptides; D021382:Protein Sorting Signals; C073768:somatotropin releasing factor 40, Leu(27)-; D000072235:Mating Factor; D013007:Growth Hormone-Releasing Hormone