PMID- 15921672
OWN - NLM
STAT- MEDLINE
DCOM- 20051215
LR  - 20131121
IS  - 0009-8981 (Print)
IS  - 0009-8981 (Linking)
VI  - 358
IP  - 1-2
DP  - 2005 Aug
TI  - Autoactivation profiles of calcium-dependent matrix metalloproteinase-2 and -9 in 
      inflammatory synovial fluid: effect of pyrophosphate and bisphosphonates.
PG  - 182-91
AB  - BACKGROUND: The presence of matrix metalloproteinase-2 and -9 (MMP-2, MMP-9), 
      gelatinase A and B, in synovial fluid is typical in inflammatory connective 
      tissue diseases especially rheumatoid arthritis (RA). Because MMPs are 
      synthesized as latent proforms, a pathophysiologic understanding of MMP 
      regulation has focused on mechanisms of activation that remain to date largely 
      unresolved. METHODS: Synovial fluid was collected by aseptic aspiration from RA 
      patients and incubated with and without physiologic levels of calcium and other 
      modifiers (pyrophosphate, bisphosphonates, and the tissue inhibitors of MMPs 
      (TIMPs), under conditions that activate MMPs. MMP-2 and -9 were then 
      characterized by substrate gel electrophoresis (gelatin zymography) to resolve 
      both latent and activated 'partially proteolyzed' forms. RESULTS: Gelatin 
      zymography revealed that RA synovial fluid contained latent neutrophil MMP-9 (92, 
      130, 225 kDa) and fibroblast MMP-2 (72 kDa). A small amount of activated MMP-2 
      (64 kDa) was also noted. Incubation of synovial fluid without calcium resulted in 
      MMP-9 activation to 87, 116, and 209 kDa forms. MMP-9 activation was, however, 
      substantially delayed in the presence of physiologic calcium (2.5 mmol/l). MMP-2 
      did not demonstrate any appreciable activation with or without physiologic 
      calcium. MMP-9 activation likely occurred via an autoactivation mechanism since 
      it was susceptible to inhibition by the tissue inhibitor of MMP-9 (TIMP-1). 
      Pyrophosphate and bisphosphonates (alendronate and risedronate) were ineffective 
      in blocking synovial fluid MMP-9 autoactivation. Some early MMP-9 activation was 
      noted with alendronate despite the presence of physiologic calcium. DISCUSSION: 
      Although RA synovial fluid contained abundant MMP-2 and MMP-9, only MMP-9 
      underwent autoactivation to lower molecular weight forms. MMP-9 was transiently 
      stable in the presence of physiologic calcium concentration, whereas 
      autoactivation was more pronounced without exogenous calcium. The apparent lack 
      of MMP-2 autoactivation with or without calcium, likely resulted from the 
      coexistence of its bound endogenous inhibitor, TIMP-2. The role of differential 
      autoactivation of MMPs activity in inflammatory arthritic disease is discussed.
FAU - Makowski, Gregory S
AU  - Makowski GS
AD  - Department of Laboratory Medicine, School of Medicine, University of Connecticut 
      Health Center, MC-2235, 263 Farmington Avenue, Farmington, CT 06030-2235, United 
      States. makowski@nso1.uchc.edu
FAU - Ramsby, Melinda L
AU  - Ramsby ML
LA  - eng
PT  - Journal Article
PL  - Netherlands
TA  - Clin Chim Acta
JT  - Clinica chimica acta; international journal of clinical chemistry
JID - 1302422
RN  - 0 (Diphosphates)
RN  - 0 (Diphosphonates)
RN  - EC 3.4.24.24 (Matrix Metalloproteinase 2)
RN  - EC 3.4.24.35 (Matrix Metalloproteinase 9)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Aged
MH  - Arthritis, Rheumatoid/*enzymology
MH  - Calcium/chemistry/metabolism/pharmacology
MH  - Diphosphates/*pharmacology/therapeutic use
MH  - Diphosphonates/*pharmacology/therapeutic use
MH  - Enzyme Activation/drug effects
MH  - Female
MH  - Humans
MH  - Inflammation/enzymology
MH  - Male
MH  - Matrix Metalloproteinase 2/chemistry/drug effects/*metabolism
MH  - Matrix Metalloproteinase 9/chemistry/drug effects/*metabolism
MH  - Middle Aged
MH  - Molecular Structure
MH  - Synovial Fluid/*enzymology
EDAT- 2005/06/01 09:00
MHDA- 2005/12/16 09:00
CRDT- 2005/06/01 09:00
PHST- 2004/12/15 00:00 [received]
PHST- 2005/03/01 00:00 [revised]
PHST- 2005/03/01 00:00 [accepted]
PHST- 2005/06/01 09:00 [pubmed]
PHST- 2005/12/16 09:00 [medline]
PHST- 2005/06/01 09:00 [entrez]
AID - S0009-8981(05)00152-X [pii]
AID - 10.1016/j.cccn.2005.03.012 [doi]
PST - ppublish
SO  - Clin Chim Acta. 2005 Aug;358(1-2):182-91. doi: 10.1016/j.cccn.2005.03.012.