PMID- 10318829
OWN - NLM
STAT- MEDLINE
DCOM- 19990617
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 20
DP  - 1999 May 14
TI  - Targeting proteins to the lumen of endoplasmic reticulum using N-terminal domains
      of 11beta-hydroxysteroid dehydrogenase and the 50-kDa esterase.
PG  - 14122-9
AB  - Previous studies identified two intrinsic endoplasmic reticulum (ER) proteins,
      11beta-hydroxysteroid dehydrogenase, isozyme 1 (11beta-HSD) and the 50-kDa
      esterase (E3), sharing some amino acid sequence motifs in their N-terminal
      transmembrane (TM) domains. Both are type II membrane proteins with the C
      terminus projecting into the lumen of the ER. This finding implied that the
      N-terminal TM domains of 11beta-HSD and E3 may constitute a lumenal targeting
      signal (LTS). To investigate this hypothesis we created chimeric fusions using
      the putative targeting sequences and the reporter gene, Aequorea victoria green
      fluorescent protein. Transfected COS cells expressing LTS-green fluorescent
      protein chimeras were examined by fluorescent microscopy and electron microscopic
      immunogold labeling. The orientation of expressed chimeras was established by
      immunocytofluorescent staining of selectively permeabilized COS cells. In
      addition, protease protection assays of membranes in the presence and absence of 
      detergents was used to confirm lumenal or the cytosolic orientation of the
      constructed chimeras. To investigate the general applicability of the proposed
      LTS, we fused the N terminus of E3 to the N terminus of the NADH-cytochrome b5
      reductase lacking the myristoyl group and N-terminal 30-residue membrane anchor. 
      The orientation of the cytochrome b5 reductase was reversed, from cytosolic to
      lumenal projection of the active domain. These observations establish that an
      amino acid sequence consisting of short basic or neutral residues at the N
      terminus, followed by a specific array of hydrophobic residues terminating with
      acidic residues, is sufficient for lumenal targeting of single-pass proteins that
      are structurally and functionally unrelated.
FAU - Mziaut, H
AU  - Mziaut H
AD  - Department of Biochemistry, University of Connecticut Health Center, Farmington, 
      Connecticut 06030-3305, USA.
FAU - Korza, G
AU  - Korza G
FAU - Hand, A R
AU  - Hand AR
FAU - Gerard, C
AU  - Gerard C
FAU - Ozols, J
AU  - Ozols J
LA  - eng
GR  - R01 GM-26351/GM/NIGMS NIH HHS/United States
PT  - Journal Article
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  - EC 1.1.- (Hydroxysteroid Dehydrogenases)
RN  - EC 1.1.1.146 (11-beta-Hydroxysteroid Dehydrogenases)
RN  - EC 1.6.2.- (Cytochrome Reductases)
RN  - EC 1.6.2.2 (Cytochrome-B(5) Reductase)
RN  - EC 3.1.1.- (Carboxylic Ester Hydrolases)
RN  - EC 3.1.1.1 (Carboxylesterase)
RN  - EC 3.4.21.64 (Endopeptidase K)
RN  - W980KJ009P (Corticosterone)
SB  - IM
MH  - 11-beta-Hydroxysteroid Dehydrogenases
MH  - Amino Acid Sequence
MH  - Animals
MH  - COS Cells
MH  - Carboxylesterase
MH  - Carboxylic Ester Hydrolases/*metabolism
MH  - Cell Membrane/metabolism
MH  - Corticosterone/metabolism
MH  - Cytochrome Reductases/metabolism
MH  - Cytochrome-B(5) Reductase
MH  - Endopeptidase K/metabolism
MH  - Endoplasmic Reticulum/*enzymology
MH  - Hydroxysteroid Dehydrogenases/*metabolism
MH  - Microscopy, Electron
MH  - Molecular Sequence Data
MH  - Rats
EDAT- 1999/05/13 00:00
MHDA- 1999/05/13 00:01
CRDT- 1999/05/13 00:00
PHST- 1999/05/13 00:00 [pubmed]
PHST- 1999/05/13 00:01 [medline]
PHST- 1999/05/13 00:00 [entrez]
AID - 10.1074/jbc.274.20.14122 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 May 14;274(20):14122-9. doi: 10.1074/jbc.274.20.14122.