PMID- 10235115
OWN - NLM
STAT- MEDLINE
DCOM- 19990528
LR  - 20061115
IS  - 1044-5498 (Print)
IS  - 1044-5498 (Linking)
VI  - 18
IP  - 4
DP  - 1999 Apr
TI  - Confocal microscopy reveals thimet oligopeptidase (EC 3.4.24.15) and neurolysin
      (EC 3.4.24.16) in the classical secretory pathway.
PG  - 323-31
AB  - Thimet oligopeptidase (EC 3.4.24.15; EP24.15) and neurolysin (EC 3.4.24.16;
      EP24.16) are closely related enzymes involved in the metabolic inactivation of
      bioactive peptides. Both of these enzymes were previously shown to be secreted
      from a variety of cell types, although their primary sequence lacks a signal
      peptide. To investigate the mechanisms responsible for this secretion, we
      examined by confocal microscopy the subcellular localization of these two enzymes
      in the neuroendocrine cell line AtT20. Both EP24.15 and EP24.16 were found by
      immunohistochemistry to be abundantly expressed in AtT20 cells. Western blotting 
      experiments confirmed that the immunoreactivity detected in the soma of these
      cells corresponded to previously cloned isoforms of the enzymes. At the
      subcellular level, both enzymes colocalized extensively with the integral
      trans-Golgi network protein, syntaxin-6, in the juxtanuclear region. In addition,
      both EP24.15 and EP24.16 were found within small vesicular organelles distributed
      throughout the cell body. Some, but not all, of these organelles also stained
      positively for ACTH. These results demonstrate that both EP24.15 and EP24.16 are 
      present within the classical secretory pathway. Their colocalization with ACTH
      further suggests that they may be targeted to the regulated secretory pathway,
      even in the absence of a signal peptide.
FAU - Garrido, P A
AU  - Garrido PA
AD  - Department of Histology and Embryology, Biomedical Science Institute, University 
      of Sao Paulo, Brazil.
FAU - Vandenbulcke, F
AU  - Vandenbulcke F
FAU - Ramjaun, A R
AU  - Ramjaun AR
FAU - Vincent, B
AU  - Vincent B
FAU - Checler, F
AU  - Checler F
FAU - Ferro, E
AU  - Ferro E
FAU - Beaudet, A
AU  - Beaudet A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - DNA Cell Biol
JT  - DNA and cell biology
JID - 9004522
RN  - EC 3.4.24.- (Metalloendopeptidases)
RN  - EC 3.4.24.15 (thimet oligopeptidase)
RN  - EC 3.4.24.16 (neurolysin)
SB  - IM
MH  - Animals
MH  - Blotting, Western
MH  - Metalloendopeptidases/*metabolism
MH  - Microscopy, Confocal/*methods
MH  - Rabbits
EDAT- 1999/05/11 00:00
MHDA- 1999/05/11 00:01
CRDT- 1999/05/11 00:00
PHST- 1999/05/11 00:00 [pubmed]
PHST- 1999/05/11 00:01 [medline]
PHST- 1999/05/11 00:00 [entrez]
AID - 10.1089/104454999315385 [doi]
PST - ppublish
SO  - DNA Cell Biol. 1999 Apr;18(4):323-31. doi: 10.1089/104454999315385.