Human betacellulin, a member of the EGF family dominantly expressed in pancreas and small intestine, is fully active in a monomeric form.
Betacellulin (BTC) was found to be expressed mainly in human pancreas and small intestine. This finding suggests that BTC possesses some specific function distinguished from the other members of epidermal growth factor (EGF) family. To clarify this function, the released form of human BTC has been expressed in E.coli, purified, and characterized. The recombinant human BTC was produced as an inclusion body. This material was dissolved in guanidine-HCl under reducing conditions, refolded, and purified through sequential liquid chromatography. Purified BTC was electrophoresed under reducing conditions and a molecular size of 18 kDa was determined, which is the supposed size of a dimer of the peptide. However, chemical analysis failed to show a covalently linked dimer. The molecular mass of BTC analyzed by mass spectrometry revealed it to be 9 kDa, which is consistent with theoretical value for a monomer. Recombinant BTC showed growth promoting activity for mouse fibroblasts and rat aortic smooth muscle cells which was equivalent to EGF On the other hand, BTC was found to exhibit a growth inhibitory effect on the cells overexpressing EGF receptor.
Seno
M
M
Department of Bioengineering Science, Faculty of Engineering, Okayama University, Japan. senom@cc.okayama-u.ac.jp
Tada
H
H
Kosaka
M
M
Sasada
R
R
Igarashi
K
K
Shing
Y
Y
Folkman
J
J
Ueda
M
M
Yamada
H
H
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D066259:Betacellulin; D006133:Growth Substances / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D007421:Intestine, Small / Q000378:metabolism*; D013058:Mass Spectrometry; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010179:Pancreas / Q000378:metabolism*; D011487:Protein Conformation; D051381:Rats; D013329:Structure-Activity Relationship
C587851:BTC protein, human; D066259:Betacellulin; C587852:Btc protein, mouse; C587853:Btc protein, rat; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins