PMID- 10428864
OWN - NLM
STAT- MEDLINE
DCOM- 19990902
LR  - 20200111
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 32
DP  - 1999 Aug 6
TI  - Autocatalytic processing of site-1 protease removes propeptide and permits
      cleavage of sterol regulatory element-binding proteins.
PG  - 22795-804
AB  - Site-1 protease (S1P) is a subtilisin-related protease that cleaves sterol
      regulatory element-binding proteins (SREBPs) in the endoplasmic reticulum lumen, 
      thereby initiating a process by which the transcriptionally active NH(2)-terminal
      fragments of SREBPs are released from membranes. In the current experiments, we
      transfected cDNAs encoding epitope-tagged hamster S1P into HEK-293 cells or
      mutant hamster cells that lack S1P. Protease protection assays showed that the
      bulk of S1P is in the endoplasmic reticulum lumen, anchored by a COOH-terminal
      membrane-spanning segment. Cleavage of the NH(2)-terminal signal sequence of S1P 
      generates S1P-A (amino acids 23-1052), which is inactive. The protein is
      self-activated by an intramolecular cleavage at Site-B, generating S1P-B (amino
      acids 138-1052) and liberating a 115-amino acid propeptide that is secreted
      intact into the medium. The sequence at Site-B is RSLK, which differs from the
      RSVL sequence at the cleavage site in SREBP-2. S1P-B is further cleaved at an
      internal RRLL sequence to yield S1P-C (amino acids 187-1052). Mutational analysis
      suggests that S1P-B and S1P-C are both active in cleaving SREBP-2 in a fashion
      that requires SREBP cleavage-activating protein. The activity of S1P-C may be
      short-lived because it appears to be transported to the Golgi, a site at which
      SREBP-2 cleavage may not normally occur. These data provide the initial
      description of the processing of a subtilisin-related protease that controls the 
      level of cholesterol in blood and cells. In an accompanying paper (Cheng, D.,
      Espenshade, P. J., Slaughter, C. A., Jaen, J. C., Brown, M. S., and Goldstein, J.
      L. (1999), J. Biol. Chem., 274, 22805-22812), we develop an in vitro assay to
      characterize the activity of purified recombinant S1P.
FAU - Espenshade, P J
AU  - Espenshade PJ
AD  - Department of Molecular Genetics, University of Texas Southwestern Medical
      Center, Dallas, Texas 75235, USA.
FAU - Cheng, D
AU  - Cheng D
FAU - Goldstein, J L
AU  - Goldstein JL
FAU - Brown, M S
AU  - Brown MS
LA  - eng
GR  - HL20948/HL/NHLBI NIH HHS/United States
GR  - HL09993/HL/NHLBI NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Hydroxycholesterols)
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (Membrane Proteins)
RN  - 0 (Protein Precursors)
RN  - 0 (Proto-Oncogene Proteins c-myc)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (SREBP cleavage-activating protein)
RN  - 0 (Sterol Regulatory Element Binding Protein 2)
RN  - 0 (Transcription Factors)
RN  - 767JTD2N31 (25-hydroxycholesterol)
RN  - 97C5T2UQ7J (Cholesterol)
RN  - EC 3.4.21.- (Proprotein Convertases)
RN  - EC 3.4.21.- (Serine Endopeptidases)
RN  - EC 3.4.21.112 (membrane-bound transcription factor peptidase, site 1)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Cholesterol/pharmacology
MH  - Cricetinae
MH  - DNA-Binding Proteins/*metabolism
MH  - Enzyme Activation
MH  - Hydroxycholesterols/pharmacology
MH  - Intracellular Signaling Peptides and Proteins
MH  - Membrane Proteins/metabolism
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Mutation
MH  - Precipitin Tests
MH  - *Proprotein Convertases
MH  - Protein Precursors/genetics/*metabolism
MH  - *Protein Processing, Post-Translational/drug effects
MH  - Proto-Oncogene Proteins c-myc/genetics
MH  - Recombinant Fusion Proteins/metabolism
MH  - Sequence Homology, Amino Acid
MH  - Serine Endopeptidases/genetics/*metabolism
MH  - Species Specificity
MH  - Sterol Regulatory Element Binding Protein 2
MH  - Transcription Factors/*metabolism
EDAT- 1999/07/31 00:00
MHDA- 1999/07/31 00:01
CRDT- 1999/07/31 00:00
PHST- 1999/07/31 00:00 [pubmed]
PHST- 1999/07/31 00:01 [medline]
PHST- 1999/07/31 00:00 [entrez]
AID - 10.1074/jbc.274.32.22795 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Aug 6;274(32):22795-804. doi: 10.1074/jbc.274.32.22795.