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 ETC possesses some spe cific function distinguished from the other members of epidermal growt h factor (EGF) family. To clarify this function, the released form of human ETC has been expressed in E.coli, purified, and characterized. T he recombinant human ETC was produced as an inclusion body. This mater ial was dissolved in guanidine-HCl under reducing conditions, refolded , and purified through sequential liquid chromatography. Purified ETC was electrophoresed under reducing conditions and a molecular size of 18 kDa was determined, which is the supposed size of a dimer of the pe ptide. However, chemical analysis failed to show a covalently linked d imer. The molecular mass of ETC analyzed by mass spectrometry revealed it to be 9 kDa, which is consistent with theoretical value for a mono mer. Recombinant ETC showed growth promoting activity for mouse fibrob lasts and rat aortic smooth muscle cells which was equivalent to EGF. On the other hand, ETC was found to exhibit a growth inhibitory effect on the cells overexpressing EGF receptor.