Structure, function, and control of neutrophil proteinases

doi:10.1016/S0002-9343(88)80068-8

The American Journal of Medicine

Volume 84, Supplement 6, 24 June 1988, Pages 37-42

https://doi.org/10.1016/S0002-9343(88)80068-8 Get rights and content

Elastase and cathepsin G are two of the major enzymes present in and secreted by human neutrophils. These proteinases can rapidly degrade connective tissue proteins. However, they also may be involved in other processes, including the activation or inactivation of protein hormones and the inactivation of plasma proteinase inhibitors. Neutrophil elastase has been implicated in the development of pulmonary emphysema, although a function for cathepsin G has not yet been elucidated. Both enzymes are normally tightly controlled by plasma proteinase inhibitors. However, this proteinase-proteinase inhibitor balance can be perturbed in favor of free enzyme by several methods, with resulting tissue damage. The use of inhibitors from several sources should be helpful in augmenting natural levels so that homeostasis can be maintained.

References (40)

MoriharaK et al.
J Biol Chem
(1965)
BernickJJ et al.
Comp Biochem Physiol
(1976)
LewisUJ et al.
J Biol Chem
(1956)
BoudierC et al.
Biochem Med
(1980)
LestienneP et al.
J Biol Chem
(1980)
BeattyK et al.
J Biol Chem
(1980)
JohnsonD et al.
J Biol Chem
(1978)
JohnsonD et al.
J Biol Chem
(1979)
MathesonNR et al.
Biochem Biophys Res Commun
(1979)
SalvesenGS et al.
J Biol Chem
(1985)

BrowerMS et al.

J Biol Chem

(1982)

RoughleyP

Biochem J

(1977)

RoughleyP et al.

Biochem J

(1977)

Del MarEG et al.

Biochemistry

(1980)

ReillyCF et al.

Biochim Biophys Acta

(1980)

BaughR et al.

Biochemistry

(1976)

SinhaS et al.

BiethJ

Frontiers Matrix Biol

(1978)

BodeW et al.

EMBO J

(1986)

ReillyCF et al.

J Biol Chem

(1982)

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Citation Excerpt :
Particularly, human neutrophil elastase (HNE) has been the subject of intensive studies because of its ability to destroy tissues. This potent proteolytic enzyme is capable of degrading or activating various proteins, including extracellular matrix proteins, receptors and cytokines [1,4,5]. HNE is involved in skin aging and lung inflammatory diseases [6–10].
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: This research was sponsored in part by grants from the National Lung Institute and The Council for Tobacco Research U.S.A

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Structure, function, and control of neutrophil proteinases

J Biol Chem

Comp Biochem Physiol

J Biol Chem

Biochem Med

J Biol Chem

J Biol Chem

J Biol Chem

J Biol Chem

Biochem Biophys Res Commun

J Biol Chem

J Biol Chem

Biochem J

Biochem J

Biochemistry

Biochim Biophys Acta

Biochemistry

Frontiers Matrix Biol

EMBO J

J Biol Chem