A mutation of furin causes the lack of precursor-processing activity in human colon carcinoma LoVo cells.
Furin has been proposed to be the endoprotease responsible for precursor cleavage at Arg-X-Lys/Arg-Arg (RXK/RR) sites within the constitutive secretory pathway. However, there was a possibility that other protease(s) is involved in this cleavage. We here characterized furin in human colon carcinoma LoVo cells, since these cells lacked the endogenous processing activity toward RXK/RR sites and recovered the activity by transfection of furin cDNA. Furin cDNA cloned from LoVo cells had one nucleotide deletion in the region covering the homo B domain which is essential for the endoproteolytic activity. LoVo cells transfected with a furin construct with the mutation showed no activity. Based on these data, we conclude that furin is the endoprotease that is involved in the precursor cleavage at RXK/RR sites within the constitutive secretory pathway.
Takahashi
S
S
Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.
Kasai
K
K
Hatsuzawa
K
K
Kitamura
N
N
Misumi
Y
Y
Ikehara
Y
Y
Murakami
K
K
Nakayama
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D003001:Cloning, Molecular; D003110:Colonic Neoplasms; D006224:Cricetinae; D045683:Furin; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009838:Oligodeoxyribonucleotides; D011061:Poly A / Q000302:isolation & purification / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011499:Protein Processing, Post-Translational; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D012333:RNA, Messenger; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity; D013381:Subtilisins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured
D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D013381:Subtilisins; D045683:Furin