PMID- 10196205
OWN - NLM
STAT- MEDLINE
DCOM- 19990517
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 16
DP  - 1999 Apr 16
TI  - Identification of a nuclear targeting domain in the insertion between helices C
      and D in protease inhibitor-10.
PG  - 11194-8
AB  - Protease inhibitor 10 (PI-10), an intracellular ovalbumin-serpin, contains a
      series of basic amino acids in the loop between helices C and D that exhibit
      homology to known nuclear targeting signals. Transfection of HeLa cells with
      plasmids encoding enhanced green fluorescent protein (EGFP) coupled to PI-10
      revealed an intense fluorescence of the nucleus. Immunoblotting demonstrated a
      single Mr 80,000 EGFP.PI-10 complex in isolated nuclei. Mutation of four basic
      amino acids in the interhelical loop to alanines (i.e. K74A, K75A, R76A, K77A)
      resulted in the fluorescent complex being confined to the cytoplasm. Further
      evidence for a nuclear targeting signal in this region was provided by
      localization of the fluorescent label to the nucleus in cells transfected with a 
      plasmid encoding EGFP fused to the 25 amino acids comprising the interhelical
      loop of PI-10 (i.e. Arg-63 to Glu-87), whereas a cytoplasmic distribution was
      noted for the construct encoding EGFP coupled to the mutated interhelical loop.
      These data raise the possibility that PI-10 may play a role in regulating
      protease activity within the nucleus, a property unique in the field of serpin
      biology.
FAU - Chuang, T L
AU  - Chuang TL
AD  - Department of Vascular Biology, The Scripps Research Institute, La Jolla,
      California 92037, USA.
FAU - Schleef, R R
AU  - Schleef RR
LA  - eng
GR  - HL45954/HL/NHLBI NIH HHS/United States
GR  - HL49563/HL/NHLBI 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  - 0 (DNA Primers)
RN  - 0 (Luminescent Proteins)
RN  - 0 (Protein Sorting Signals)
RN  - 0 (SERPINB10 protein, human)
RN  - 0 (Serpins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Amino Acid Sequence
MH  - Base Sequence
MH  - Blotting, Western
MH  - Cell Nucleus/*metabolism
MH  - DNA Primers
MH  - Fluorescence
MH  - Genes, Reporter
MH  - Green Fluorescent Proteins
MH  - HeLa Cells
MH  - Humans
MH  - Luminescent Proteins/genetics
MH  - Molecular Sequence Data
MH  - Plasmids
MH  - Protein Sorting Signals/*metabolism
MH  - Serpins/chemistry/*metabolism
EDAT- 1999/04/10 00:00
MHDA- 1999/04/10 00:01
CRDT- 1999/04/10 00:00
PHST- 1999/04/10 00:00 [pubmed]
PHST- 1999/04/10 00:01 [medline]
PHST- 1999/04/10 00:00 [entrez]
AID - 10.1074/jbc.274.16.11194 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Apr 16;274(16):11194-8. doi: 10.1074/jbc.274.16.11194.