PMID- 10430032
OWN - NLM
STAT- MEDLINE
DCOM- 19990831
LR  - 20091119
IS  - 1431-6730 (Print)
IS  - 1431-6730 (Linking)
VI  - 380
IP  - 6
DP  - 1999 Jun
TI  - Localization of rat cathepsin K in osteoclasts and resorption pits: inhibition of
      bone resorption and cathepsin K-activity by peptidyl vinyl sulfones.
PG  - 679-87
AB  - We have localized cathepsin K in rat osteoclasts and within exposed resorption
      pits by immuno-fluorescence microscopy. Intracellular staining using an antibody 
      raised against recombinant mouse cathepsin K was vesicular and uniformly
      distributed throughout the cell. Confocal microscopy analysis did not reveal an
      accumulation of cathepsin K containing vesicles opposing the ruffled border and
      the resorption lacuna. Exposed resorption pits exhibited a uniform distribution
      of cathepsin K, and no differences were observed between the edges and the
      centers of the pits. The immunostaining of resorption pits with anti-cathepsin K 
      antibodies demonstrates that the protease is secreted into the sub-osteoclastic
      compartment. Cathepsin K-specific inhibition using peptidyl vinyl sulfones as
      selective cysteine protease inactivators reduced bone resorption by 80% in a
      dose-dependent manner at sub-micromolar concentrations. No reduction of bone
      resorption was observed at those low concentrations using a potent cathepsin L,
      S, B-specific inhibitor. That the inhibition of bone resorption can be attributed
      to cathepsin K-like protease inhibition was corroborated by the selective
      inhibition of the osteoclastic Z-Gly-Pro-Arg-MbetaNA hydrolyzing activity by the 
      cathepsin K, L, S, B-inhibitor, but not by the cathepsin L, B, and S inhibitor.
      Z-Gly-Pro-Arg-MbetaNA is efficiently hydrolyzed by cathepsin K but only poorly by
      cathepsins L, S, and B. On the contrary, the intracellular hydrolysis of the
      cathepsin B-specific substrate, Z-Arg-Arg-MbetaNA, was prevented by both types of
      inhibitors. The identification of cathepsin K in resorption pits and the
      inhibition of bone resorption and intracellular cathepsin K activity by selective
      vinyl sulfone inhibitors indicate the critical role of the protease in
      osteoclastic bone resorption.
FAU - Xia, L
AU  - Xia L
AD  - Mount Sinai School of Medicine, Department of Human Genetics, New York, NY 10029,
      USA.
FAU - Kilb, J
AU  - Kilb J
FAU - Wex, H
AU  - Wex H
FAU - Li, Z
AU  - Li Z
FAU - Lipyansky, A
AU  - Lipyansky A
FAU - Breuil, V
AU  - Breuil V
FAU - Stein, L
AU  - Stein L
FAU - Palmer, J T
AU  - Palmer JT
FAU - Dempster, D W
AU  - Dempster DW
FAU - Bromme, D
AU  - Bromme D
LA  - eng
GR  - 1 S10 RR0 9145-01/RR/NCRR NIH HHS/United States
GR  - AR 39191/AR/NIAMS NIH HHS/United States
GR  - AR 41331/AR/NIAMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Germany
TA  - Biol Chem
JT  - Biological chemistry
JID - 9700112
RN  - 0 (Cysteine Proteinase Inhibitors)
RN  - 0 (DNA Primers)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Sulfones)
RN  - EC 3.4.- (Cathepsins)
RN  - EC 3.4.22.38 (CTSK protein, human)
RN  - EC 3.4.22.38 (Cathepsin K)
RN  - EC 3.4.22.38 (Ctsk protein, mouse)
RN  - EC 3.4.22.38 (Ctsk protein, rat)
SB  - IM
SB  - S
MH  - Animals
MH  - Base Sequence
MH  - Bone Resorption/*prevention & control
MH  - Cathepsin K
MH  - Cathepsins/antagonists & inhibitors/*metabolism
MH  - Cell Compartmentation
MH  - Cells, Cultured
MH  - Cysteine Proteinase Inhibitors/pharmacology
MH  - DNA Primers
MH  - Immunohistochemistry
MH  - Mice
MH  - Osteoclasts/*enzymology
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Recombinant Proteins/antagonists & inhibitors/metabolism
MH  - Sulfones/*pharmacology
EDAT- 1999/08/03 00:00
MHDA- 1999/08/03 00:01
CRDT- 1999/08/03 00:00
PHST- 1999/08/03 00:00 [pubmed]
PHST- 1999/08/03 00:01 [medline]
PHST- 1999/08/03 00:00 [entrez]
AID - 10.1515/BC.1999.084 [doi]
PST - ppublish
SO  - Biol Chem. 1999 Jun;380(6):679-87. doi: 10.1515/BC.1999.084.