Functional expression of furin demonstrating its intracellular localization and endoprotease activity for processing of proalbumin and complement pro-C3.
We have cloned a rat cDNA encoding furin which is structurally related to yeast Kex2 protease. Products of 88 and 94 kDa were obtained by in vitro transcription/translation of the cDNA in the absence and presence of microsomes. When the cDNA was transfected into COS-1 cells, furin was expressed as a major glycosylated form of 94 kDa, accompanied by a minor proteolytic form of 86 kDa, and found to be localized in the Golgi complex. Proalbumin and complement pro-C3 are intracellularly processed into their mature forms by cleavage at the dibasic residues Arg-Arg, a common cleavage signal found in many pro-type precursors. In cells transfected with the cDNA of C3 or albumin alone, only about half of each proform expressed was processed by an endogenous activity of the cells. When furin was coexpressed, the proforms of both C3 and albumin were completely processed into their mature forms. In addition, co-expression of rat alpha 1-protease inhibitor mutant (Met352----Arg) resulted in inhibition of the endogenous and exogenous processing activities, as observed for the naturally occurring mutant Pittsburgh which has been identified as a specific inhibitor for the processing enzyme. Taken together, these results indicate that furin is an endoprotease localized to the Golgi complex and capable of processing proalbumin and pro-C3 into the mature forms.
Misumi
Y
Y
Department of Biochemistry, Fukuoka University School of Medicine, Japan.
Oda
K
K
Fujiwara
T
T
Takami
N
N
Tashiro
K
K
Ikehara
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003176:Complement C3 / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D045683:Furin; D008969:Molecular Sequence Data; D011228:Prealbumin / Q000378:metabolism*; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D051381:Rats; D013381:Subtilisins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection
D003176:Complement C3; D011228:Prealbumin; D011498:Protein Precursors; D013381:Subtilisins; D045683:Furin