Characterization of plasma gelsolin as a substrate for matrix metalloproteinases.
We previously showed that plasma gelsolin, a major component of the extracellular actin scavenging system, is an matrix metalloproteinase (MMP)-14 substrate. Here we confirmed that plasma gelsolin is cleaved by MMP-14 at the plasma level, and found that it was most efficiently digested by MMP-3 followed by MMP-2, MMP-1, MMP-14, and MMP-9, in that order. Plasma gelsolin (90 kDa) was cut into several fragments of 43-48 kDa by MMP-3. The MMP-3 cleavage sites in plasma gelsolin were determined by labeling the C termini generated by in-gel digestion with 50% H2 18O combined with peptide mass mapping, and sequencing of the N-terminal amino acids. Plasma gelsolin was cleaved at Asn416-Val417, Ser51-Met52, and Ala435-Gln436. Proteolytic cleavage by MMP-3 resulted in considerable loss of its actin filament-depolymerizing activity. This suggests that MMPs weaken the extracellular actin-scavenging system by cleaving plasma gelsolin and may, therefore, be involved in pathological conditions induced by extracellular actin, such as endothelial injury, respiratory distress syndrome, hepatic necrosis, and septic shock.
Park
Sung-Min
SM
Department of Biochemistry, College of Science, Yonsei University, Seoul, Korea.
Hwang
In Kwan
IK
Kim
Se Yeon
SY
Lee
Seo-Jin
SJ
Park
Kang-Sik
KS
Lee
Seung-Taek
ST
D000199:Actins / Q000378:metabolism; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D018260:Gelsolin / Q000097:blood*; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000494:pharmacology; D020778:Matrix Metalloproteinase 2 / Q000494:pharmacology; D019278:Matrix Metalloproteinase 3 / Q000494:pharmacology; D020780:Matrix Metalloproteinase 9 / Q000494:pharmacology; D020782:Matrix Metalloproteinases / Q000378:metabolism* / Q000494:pharmacology; D053504:Matrix Metalloproteinases, Membrane-Associated; D010449:Peptide Mapping; D011994:Recombinant Proteins / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity
D000199:Actins; D018260:Gelsolin; D011994:Recombinant Proteins; D020782:Matrix Metalloproteinases; D053504:Matrix Metalloproteinases, Membrane-Associated; D019278:Matrix Metalloproteinase 3; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D020781:Matrix Metalloproteinase 1