Elastase

The Uses of Elastase

Elastase is used in combination with other proteases for protein analysis by mass spectrometry. Elastase, is used in tissue dissociation because elastin is found in highest concentrations in the elastic fibers of connective tissues, elastase is frequently used to dissociate tissues that contain extensive intercellular fiber networks. It is usually used with other enzymes such as collagenase, trypsin, and chymotrypsin, membrane protein solubilization, protein sequence studies.

The Uses of Elastase

Elastase from porcine pancreas has been used in a study to identify a novel bone/calcium metabolism-regulating factor in porcine pancreas. Elastase from porcine pancreas has also been used in a study to investigate the effect of a specific synthetic inhibitor of neutrophil elastase (ONO-5046) on the course of acute hemorrhagic pancreatitis in dogs.

The Uses of Elastase

Elastase from porcine pancreas has been used:

• to treat vero cells?to study its effects on syncytium formation
• as a positive control in protease assays
• as a component in RPMI 1640 to isolate human aortic smooth muscle cells (HASMCs) from the aortic tissue

General Description

Elastase is a single polypeptide chain of 240 amino acid residues and contains four disulfide bridges. The molecular mass is approximately 25.9kDa. The enzyme is synthesized as an inactive zymogen, proelastase, which is converted to the active form by limited proteolysis at the N-terminal by trypsin.

Biochem/physiol Actions

Elastase is a single polypeptide chain of 240 amino acid residues and contains four disulfide bridges. The molecular mass is approximately 25.9 kDa. The enzyme is synthesized as an inactive zymogen, proelastase, which is converted to the active form by limited proteolysis at the N-terminal by trypsin. It is a serine protease with broad specificity. It cleaves protein at the carboxyl side of small hydrophobic amino acids such as Ile, Gly, Ala, Ser, Val, and Leu. The enzyme also hydrolyzes amides and esters such as N-Benzoyl-L-alanine methyl ester. The pH optimum is found to be 8.0-8.5. It does not require any activator, but it is inhibited by diisopropyl fluorophosphate, phenylmethanesulfonyl fluoride, α2-macroglobulin, α1-antitrypsin, sulfonyl fluorides and p-dinitrophenyl diethylphosphate and high salt concentrations. It is extensively used in tissue and cell dissociation procedures. Elastase is effective in the isolation of Type II lung cells.