PMID- 23733191
OWN - NLM
STAT- MEDLINE
DCOM- 20130927
LR  - 20211021
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 288
IP  - 28
DP  - 2013 Jul 12
TI  - Non-destructive and selective imaging of the functionally active, pro-invasive 
      membrane type-1 matrix metalloproteinase (MT1-MMP) enzyme in cancer cells.
PG  - 20568-80
LID - 10.1074/jbc.M113.471508 [doi]
AB  - Proteolytic activity of cell surface-associated MT1-matrix metalloproteinase 
      (MMP) (MMP-14) is directly related to cell migration, invasion, and metastasis. 
      MT1-MMP is regulated as a proteinase by activation and conversion of the latent 
      proenzyme into the active enzyme, and also via inhibition by tissue inhibitors of 
      MMPs (TIMPs) and self-proteolysis. MT1-MMP is also regulated as a membrane 
      protein through its internalization and recycling. Routine immunohistochemistry, 
      flow cytometry, reverse transcription-PCR, and immunoblotting methodologies do 
      not allow quantitative imaging and assessment of the cell-surface levels of the 
      active, TIMP-free MT1-MMP enzyme. Here, we developed a fluorescent reporter 
      prototype that targets the cellular active MT1-MMP enzyme alone. The reporter 
      (MP-3653) represents a liposome tagged with a fluorochrome and functionalized 
      with a PEG chain spacer linked to an inhibitory hydroxamate warhead. Our studies 
      using the MP-3653 reporter and its inactive derivative demonstrated that MP-3653 
      can be efficiently used not only to visualize the trafficking of MT1-MMP through 
      the cell compartment, but also to quantify the femtomolar range amounts of the 
      cell surface-associated active MT1-MMP enzyme in multiple cancer cell types, 
      including breast carcinoma, fibrosarcoma, and melanoma. Thus, the levels of the 
      naturally expressed, fully functional, active cellular MT1-MMP enzyme are roughly 
      equal to 1 x 10(5) molecules/cell, whereas these levels are in a 1 x 10(6) range 
      in the cells with the enforced MT1-MMP expression. We suggest that the reporter 
      we developed will contribute to the laboratory studies of MT1-MMP and then, 
      ultimately, to the design of novel, more efficient prognostic approaches and 
      personalized cancer therapies.
FAU - Remacle, Albert G
AU  - Remacle AG
AD  - Sanford-Burnham Medical Research Institute, La Jolla, California 92037, USA.
FAU - Shiryaev, Sergey A
AU  - Shiryaev SA
FAU - Golubkov, Vladislav S
AU  - Golubkov VS
FAU - Freskos, John N
AU  - Freskos JN
FAU - Brown, Michael A
AU  - Brown MA
FAU - Karwa, Amolkumar S
AU  - Karwa AS
FAU - Naik, Arati D
AU  - Naik AD
FAU - Howard, Carol P
AU  - Howard CP
FAU - Sympson, Carolyn J
AU  - Sympson CJ
FAU - Strongin, Alex Y
AU  - Strongin AY
LA  - eng
GR  - R01DE022757/DE/NIDCR NIH HHS/United States
GR  - R01 DE022757/DE/NIDCR NIH HHS/United States
GR  - R01 CA083017/CA/NCI NIH HHS/United States
GR  - R01CA83017/CA/NCI NIH HHS/United States
GR  - R01 CA157328/CA/NCI NIH HHS/United States
GR  - R01CA157328/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20130603
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Alexa594)
RN  - 0 (Fluoresceins)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Liposomes)
RN  - 0 (Organic Chemicals)
RN  - 0 (Tissue Inhibitor of Metalloproteinase-1)
RN  - 127497-59-0 (Tissue Inhibitor of Metalloproteinase-2)
RN  - 3301-79-9 (6-carboxyfluorescein)
RN  - EC 3.4.24.80 (Matrix Metalloproteinase 14)
SB  - IM
MH  - Animals
MH  - Binding, Competitive
MH  - Blotting, Western
MH  - Cell Line
MH  - Cell Line, Tumor
MH  - Fluoresceins/chemistry
MH  - Fluorescent Dyes/chemistry
MH  - HEK293 Cells
MH  - Humans
MH  - Liposomes/chemistry/metabolism
MH  - MCF-7 Cells
MH  - Matrix Metalloproteinase 14/chemistry/genetics/*metabolism
MH  - Microscopy, Fluorescence
MH  - Molecular Imaging/*methods
MH  - Mutation
MH  - Neoplasms/*enzymology/genetics/pathology
MH  - Optical Imaging/*methods
MH  - Organic Chemicals/chemistry
MH  - Protein Binding
MH  - Tissue Inhibitor of Metalloproteinase-1/genetics/metabolism
MH  - Tissue Inhibitor of Metalloproteinase-2/genetics/metabolism
PMC - PMC3711321
OTO - NOTNLM
OT  - Breast Cancer
OT  - Cancer Biology
OT  - Drug Delivery
OT  - Enzyme Inhibitors
OT  - Hydroxamate Inhibitors
OT  - Imaging
OT  - Liposomes
OT  - MT1-MMP
OT  - Matrix Metalloproteinase (MMP)
OT  - Protease Inhibitor
EDAT- 2013/06/05 06:00
MHDA- 2013/09/28 06:00
PMCR- 2014/07/12
CRDT- 2013/06/05 06:00
PHST- 2013/06/05 06:00 [entrez]
PHST- 2013/06/05 06:00 [pubmed]
PHST- 2013/09/28 06:00 [medline]
PHST- 2014/07/12 00:00 [pmc-release]
AID - S0021-9258(20)45625-X [pii]
AID - M113.471508 [pii]
AID - 10.1074/jbc.M113.471508 [doi]
PST - ppublish
SO  - J Biol Chem. 2013 Jul 12;288(28):20568-80. doi: 10.1074/jbc.M113.471508. Epub 
      2013 Jun 3.