TY - JOUR
T1 - A Comparative Study of Fibronectin Cleavage by MMP-1, -3, -13, and -14
AU - Zhang, Xiaorong
AU - Chen, Christopher T.
AU - Bhargava, Madhu
AU - Torzilli, Peter A.
N1 - Funding Information:
The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: Support for this study was made possible by grant numbers AR46574 (P.A.T.), AR45748 (P.A.T.), and AR059203 (P.A.T.) from the National Institutes of Health, National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIH-NIAMS). Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH-NIAMS. This investigation was conducted in a facility constructed with support from Research Facilities Improvement Program grant number C06-RR12538-01 from the National Center for Research Resources, National Institutes of Health.
PY - 2012/7
Y1 - 2012/7
N2 - Fibronectin fragments are important for synovial inflammation and the progression of arthritis, and thus, identifying potential enzymatic pathways that generate these fragments is of vital importance. The objective of this study was to determine the cleavage efficiency of fibronectin by matrix metalloproteinases (MMP-1, MMP-3, MMP-13, and MMP-14). Intact human plasma fibronectin was co-incubated with activated MMPs in neutral or acidic pH for up to 24 hours at 37 °C. The size and distribution of fibronectin fragments were determined by Western blot analysis using antibodies that recognized the N-terminals of fibronectin. All MMPs were able to cleave fibronectin at neutral pH. MMP-13 and -14 had the highest efficiency followed by MMP-3 and -1. MMP-3, -13, and -14 generated 70-kDa fragments, a known pro-inflammatory peptide. Further degradation of fibronectin fragments was only found for MMP-13 and -14, generating 52-kDa, 40-kDa, 32-kDa, and 29-kDa fragments. Fibronectin fragments of similar size were also found in the articular cartilage of femoral condyles of normal bovine knee joints. At acidic pH (5.5), the activities of MMP-1 and -14 were nearly abolished, while MMP-3 had a greater efficiency than MMP-13 even though the activities of both MMPs were significantly reduced. These findings suggest that MMP-13 and -14 may play a significant role in the cleavage of fibronectin and the production of fibronectin fragments in normal and arthritic joints.
AB - Fibronectin fragments are important for synovial inflammation and the progression of arthritis, and thus, identifying potential enzymatic pathways that generate these fragments is of vital importance. The objective of this study was to determine the cleavage efficiency of fibronectin by matrix metalloproteinases (MMP-1, MMP-3, MMP-13, and MMP-14). Intact human plasma fibronectin was co-incubated with activated MMPs in neutral or acidic pH for up to 24 hours at 37 °C. The size and distribution of fibronectin fragments were determined by Western blot analysis using antibodies that recognized the N-terminals of fibronectin. All MMPs were able to cleave fibronectin at neutral pH. MMP-13 and -14 had the highest efficiency followed by MMP-3 and -1. MMP-3, -13, and -14 generated 70-kDa fragments, a known pro-inflammatory peptide. Further degradation of fibronectin fragments was only found for MMP-13 and -14, generating 52-kDa, 40-kDa, 32-kDa, and 29-kDa fragments. Fibronectin fragments of similar size were also found in the articular cartilage of femoral condyles of normal bovine knee joints. At acidic pH (5.5), the activities of MMP-1 and -14 were nearly abolished, while MMP-3 had a greater efficiency than MMP-13 even though the activities of both MMPs were significantly reduced. These findings suggest that MMP-13 and -14 may play a significant role in the cleavage of fibronectin and the production of fibronectin fragments in normal and arthritic joints.
KW - cartilage
KW - fibronectin
KW - fibronectin fragments
KW - matrix metalloproteinases
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U2 - 10.1177/1947603511435273
DO - 10.1177/1947603511435273
M3 - Article
C2 - 26069638
AN - SCOPUS:84863991406
SN - 1947-6035
VL - 3
SP - 267
EP - 277
JO - Cartilage
JF - Cartilage
IS - 3
ER -