Expression of a recombinant human growth hormone binding protein in baculovirus/insect cell system.
An eucaryotic recombinant human growth hormone binding protein (rGHBP) was expressed in baculovirus-infected insect cells and purified by affinity chromatography from culture supernatant. This mannose-rich 34-kDa protein specifically bound human growth hormone (hGH) with the same affinity (kDa = 0.42 x 10(-9) M) than the 51.5 kDa GHBP we purified and characterised from human plasma (kDa = 1.1 x 10(-9) M). A high molecular form of the rGHBP was detected by silver-stained SDS-PAGE, Western blot (mAb 263), affinity cross-linking and Western ligand blot with 125I-hGH. Reduction experiments with beta-mercaptoethanol suggested that this form involved a disulfide bound between two rGHBPs.
Cholin
S
S
Centre de Recherche sur la Croissance, Centre Hospitalier Universitaire de Purpan, Toulousel Pharmacia France.
Bieth
E
E
Molinas
C
C
Cahoreau
C
C
Perals
C
C
Cerruti
M
M
Devauchelle
G
G
Rochiccioli
P
P
Tauber
M
M
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors; D006031:Glycosylation; D019382:Human Growth Hormone; D006801:Humans; D007457:Iodine Radioisotopes; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D018411:Spodoptera / Q000166:cytology
D002352:Carrier Proteins; D007457:Iodine Radioisotopes; D011993:Recombinant Fusion Proteins; D019382:Human Growth Hormone; C051815:somatotropin-binding protein