Rat prostatic growth factors: purification and characterization of high and low molecular weight epidermal growth factors from rat dorsolateral prostate.
Growth factors which possibly participate in androgen-induced proliferation of rat prostate epithelial cells have been purified and characterized. Four distinct forms of growth factor were found in the extract of rat dorsolateral prostate. One of the factors was a member of heparin-binding growth factor (HBGF) family judging from its high affinity for heparin-Sepharose. The other three factors were capable of competing with [125I]epidermal growth factor (EGF) for the cell surface receptor, and recognized by anti-rat EGF antiserum. These EGF-like factors (EGF1-EGF3) were purified by ion-exchange chromatography, gel filtration and reverse phase HPLC. EGF1 showed microheterogeneity on chromatographic and electrophoretic separation and N-terminal sequence analysis. EGF1 showed an average molecular weight of about 35,000 on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions. These results indicated that EGF1 was a mixture of high molecular weight forms of EGF. The molecular weights of EGF2 and EGF3 were similar to that of rat submaxillary gland EGF (Mr = 5400). The amino acid sequence of EGF2 was identical with that of rat EGF except for the N- and C-terminal amino acids: aspartic acid instead of asparagine was found at the N-terminal position and C-terminal arginine was missing in EGF2. Although the N-terminal sequence of EGF3 (1-19) was identical with that of EGF2, the two factors were completely separated by gel filtration indicating a difference in the C-terminal structure. EGF1, EGF2 and EGF3 but not HBGF stimulated proliferation of primary cultured rat dorsolateral prostate epithelial cells.
Nishi
N
N
Department of Endocrinology, Kagawa Medical School, Japan.
Shimizu
C
C
Okutani
T
T
Kagawa
Y
Y
Takasuga
H
H
Suno
M
M
Wada
F
F
D000595:Amino Acid Sequence; D000818:Animals; D002455:Cell Division; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004815:Epidermal Growth Factor / Q000737:chemistry / Q000302:isolation & purification*; D004847:Epithelial Cells; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011467:Prostate / Q000033:anatomy & histology / Q000737:chemistry*; D051381:Rats; D011919:Rats, Inbred Strains
D004815:Epidermal Growth Factor