PMID- 8408037
OWN - NLM
STAT- MEDLINE
DCOM- 19931118
LR  - 20210210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 268
IP  - 29
DP  - 1993 Oct 15
TI  - Modulation of furin-mediated proprotein processing activity by site-directed 
      mutagenesis.
PG  - 21826-34
AB  - The proprotein processing activity of mutants of the subtilisin-like enzyme furin 
      was studied in transfected mammalian cells. Our studies indicate that the three 
      residues of the catalytic triad of furin, Asp46, His87, and Ser261, are critical 
      not only for substrate processing but also for maturation of furin. Furthermore, 
      evidence is provided that maturation of furin occurs through an intramolecular 
      autocatalytic process. Substitution of the asparagine residue (Asn188) of the 
      oxyanion hole by an alanine residue appears to block substrate processing but not 
      furin maturation. Analysis of carboxyl-terminal deletion mutants revealed that 
      the segment encompassing residues Glu449 to Glu469 of the "middle" domain, which 
      is more than 100 residues downstream of the predicted catalytic domain, contains 
      residues that seem to be critical for processing activity but that the more 
      carboxyl-terminal cysteine-rich region, the transmembrane region, and the 
      cytosolic tail are dispensable. Finally, we made mutants in the substrate binding 
      region of human furin and studied their ability to process von Willebrand factor 
      (pro-vWF) substrates, including wild-type pro-vWF as well as pro-vWF mutants in 
      which the P1 (vWFR-1G), P2 (vWFK-2A), or P4 (vWFR-4A) basic residue with respect 
      to the pro region cleavage site had been mutated. It is demonstrated that 
      particular negatively charged residues in or near the substrate binding region of 
      furin are critical for cleavage activity and specificity of the enzyme for 
      multiple basic residues in the substrate. Furthermore, substrate binding region 
      mutants of furin were obtained, which cleaved either the pro-vWFK-2A or 
      pro-vWFR-4A mutant of pro-vWF more efficiently than wild-type pro-vWF.
FAU - Creemers, J W
AU  - Creemers JW
AD  - Laboratory for Molecular Oncology, University of Leuven, Belgium.
FAU - Siezen, R J
AU  - Siezen RJ
FAU - Roebroek, A J
AU  - Roebroek AJ
FAU - Ayoubi, T A
AU  - Ayoubi TA
FAU - Huylebroeck, D
AU  - Huylebroeck D
FAU - Van de Ven, W J
AU  - Van de Ven WJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Amino Acids)
RN  - 0 (DNA Primers)
RN  - 0 (Protein Precursors)
RN  - 0 (von Willebrand Factor)
RN  - EC 3.4.21.- (Subtilisins)
RN  - EC 3.4.21.75 (Furin)
SB  - IM
MH  - Amino Acids/chemistry
MH  - Base Sequence
MH  - Binding Sites
MH  - Cells, Cultured
MH  - DNA Primers
MH  - Furin
MH  - Humans
MH  - Hydrolysis
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Precipitin Tests
MH  - Protein Precursors/metabolism
MH  - *Protein Processing, Post-Translational
MH  - Sequence Deletion
MH  - Subtilisins/biosynthesis/genetics/*metabolism
MH  - von Willebrand Factor/genetics/metabolism
EDAT- 1993/10/15 00:00
MHDA- 1993/10/15 00:01
CRDT- 1993/10/15 00:00
PHST- 1993/10/15 00:00 [pubmed]
PHST- 1993/10/15 00:01 [medline]
PHST- 1993/10/15 00:00 [entrez]
AID - S0021-9258(20)80616-4 [pii]
PST - ppublish
SO  - J Biol Chem. 1993 Oct 15;268(29):21826-34.