Alpha 1-antitrypsin is degraded and non-functional in chronic wounds but intact and functional in acute wounds: the inhibitor protects fibronectin from degradation by chronic wound fluid enzymes.
Fluid obtained from chronic and acute wounds were examined for the presence of fibronectin, alpha 1-antitrypsin, and proteinases capable of degrading both proteins. Immunoblot analysis of fluids from ten chronic wounds revealed that fibronectin and alpha 1-antitrypsin were degraded in nine of ten samples. In contrast, both fibronectin and alpha 1-antitrypsin were intact in acute wound fluids. The degradation of the inhibitor and fibronectin occurred in the same wound fluids, and these two events correlated perfectly. Chronic or acute wound fluid proteins were coupled to benzamidine Sepharose 6B beads and incubated with fibronectin or alpha 1-antitrypsin. Chronic wound fluid proteins degraded fibronectin in the presence of ethylenediaminetetraacetate, leupeptin, cystatin, and pepstatin but not in the presence of phenylmethylsulfonyl fluoride. Acute wound fluids and normal human serum did not contain enzymes capable of degrading fibronectin. These data suggest that serine proteinases are responsible for fibronectin degradation in chronic wound fluids. Chronic wound fluids that contained degraded alpha 1-antitrypsin also contain proteinases capable of degrading alpha 1-antitrypsin from human serum. Acute wound fluids and normal human serum did not contain enzymes capable of degrading alpha 1-antitrypsin. The inhibitor from acute wound fluids bound to one of its targets, trypsin. In contrast, the fragment(s) of alpha 1-antitrypsin from chronic wound fluids did not bind trypsin. Chronic wounds associated with degraded fibronectin and the inhibitor contained ten- to forty-fold more elastase activity than acute wounds. The degradation of fibronectin by chronic wound fluid enzymes was inhibited by alpha 1-antitrypsin in a dose-dependent manner. Collectively, these results demonstrate that there are enzymes in chronic wounds that perturb the function of alpha 1-antitrypsin and allow fibronectin degradation by uninhibited serine proteinases.
Rao
C N
CN
Department of Dermatology, Northwestern University Medical School, Chicago, Illinois 60611, USA.
Ladin
D A
DA
Liu
Y Y
YY
Chilukuri
K
K
Hou
Z Z
ZZ
Woodley
D T
DT
D000208:Acute Disease; D001940:Breast / Q000293:injuries; D002908:Chronic Disease; D004492:Edetic Acid / Q000494:pharmacology; D010450:Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism; D005122:Exudates and Transudates / Q000737:chemistry*; D005260:Female; D005353:Fibronectins / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D008297:Male; D010196:Pancreatic Elastase / Q000302:isolation & purification / Q000378:metabolism; D012867:Skin / Q000293:injuries; D014647:Varicose Ulcer / Q000378:metabolism; D014947:Wounds and Injuries / Q000378:metabolism*; D000515:alpha 1-Antitrypsin / Q000302:isolation & purification / Q000378:metabolism*
D005353:Fibronectins; D000515:alpha 1-Antitrypsin; D004492:Edetic Acid; D010450:Endopeptidases; D010196:Pancreatic Elastase