Elastase

On this page:

• Physical Properties
• Clinical Implications
• Specificity, Kinetics and Activity Determination
• Storage and Stability
• Related Products

Enzyme Commission (EC) Number: 3.4.21.37
Product Number: E8140

Synonyms:

lysosomal elastase, neutrophil elastase, polymorphonuclear leukocyte elastase, serine elastase, lysosomal elastase, granulocyte elastase

Physical Properties

Leukocyte elastase is a 29KDa serine endoprotease of the Proteinase S1 Family. It exists as a single 238 amino acid-peptide chain with four disulfide bonds. The human leukocyte enzyme shares approximately 43% sequence homology with porcine pancreatic elastase. It contains two or thee N-linked glycans of variable composition which account for its three major isoforms.^1,2,3
Isoelectric point: pI = 8.77 - 9.55³

Clinical Implications

Elastase is one of the degradative hydrolases contained in the azurophil granules of phygocytic neutrophils.³

Elevated levels of circulating elastase and/or decreased levels of endogenous inhibitors have been implicated in several inflammatory diseases including arthritis,⁴ Cyclic neutropenia,⁵ Myocardial Ischemia,⁶ Pancreatitis,⁷ Psoriasis,⁸ Pulmonary Emphysema,⁹ and endotoxic shock.¹⁰

Several inhibitors of leukocyte elastase, including elafin, and have been proposed for treatment of related diseases.

Specificity, Kinetics and Activity Determination

Unlike pancreatic elastase the leukocyte enzyme has a preferential cleavage for the carboxyl side of valine, but will also cleave to a lesser extent after alanine.^11,12,13 Natural substrates include elastin,¹⁴ cartilage proteoglycans,¹⁵ collagen types I, II, II and IV^16,17,18 and fibronectin.¹⁹

pH Optimum for activity: 8.5²⁰

Sigma’s Unit Definition: One unit will release one nanomole of p-nitrophenol per second from N-t-BOC-L-alanine p-nitrophenyl ester at pH 6.5 at 37 °C

Method: Continuous Spectrophotometric Rate Determination

In a 3.00 mL reaction mix, the final concentrations are 49 mM sodium phosphate, 0.20 mM N-t-BOC-L-alanine p-nitrophenyl ester, 2% (v/v) acetonitrile, and 1 unit of elastase.

Storage and Stability Detergents such as 0.05-0.2%Triton^® X-100 or 0.1% Brij^® 35 are often included in reaction and extraction buffers to insure solubilization and reduce adhesion to glass particularly at low ionic strength.^2,21

The enzyme retains activity after exposure to 8 M urea and retains 22% of its activity in the presence of 1% SDS. In SDS and 2-mercaptoethanol the enzyme undergoes autolysis giving rise to several low molecular weight fragments.²²

Store lyophilized powder at -20 °C
Once reconstituted, store in frozen aliquots at or below -20 °C

References

1. Sinha S, Watorek W, Karr S, Giles J, Bode W, Travis J. 1987. Primary structure of human neutrophil elastase.. Proceedings of the National Academy of Sciences. 84(8):2228-2232. https://doi.org/10.1073/pnas.84.8.2228

2. Baugh RJ, Travis J. 1976. Human leukocyte granule elastase: rapid isolation and characterization. Biochemistry. 15(4):836-841. https://doi.org/10.1021/bi00649a017

3. Barrett AJ. 1981. [44] Leukocyte Elastase.581-588. https://doi.org/10.1016/s0076-6879(81)80046-8

4. Feltelius N, Hallgren R. 1988. Circulating inhibitor bound elastase in patients with ankylosing spondylitis and rheumatoid arthritis and the influence of sulphasalazine treatment.. Annals of the Rheumatic Diseases. 47(1):10-14. https://doi.org/10.1136/ard.47.1.10

5. Horwitz M, Benson KF, Duan Z, Person RE, Wechsler J, Williams K, Albani D, Li F. 2003. Role of neutrophil elastase in bone marrow failure syndromes: molecular genetic revival of the chalone hypothesis. Current Opinion in Hematology. 10(1):49-54. https://doi.org/10.1097/00062752-200301000-00008

6. Ceriana P. 1992. Effect of Myocardial Ischaemia-reperfusion on Granulocyte Elastase Release. Anaesthesia and Intensive Care. 20(2):187-190. https://doi.org/10.1177/0310057x9202000212

7. Uehara S, Gotoh K, Handa H, Honjo K, Hiraŷama A, Sakamoto W. 1992. Changes in polymorphonuclear neutrophil-elastase in pancreatitis. Agents and actions. Supplements..(38):405-12.

8. Glinski W, Pierozynska-Dubowska M, Glinska-Ferenz M, Jab?o?ska S. 1991. Decreased specific anti-elastase activity in the uninvolved skin of patients with psoriasis. Arch Dermatol Res. 283(4):224-229. https://doi.org/10.1007/bf01106106

9. Kuraki T, Ishibashi M, Takayama M, Shiraishi M, Yoshida M. 2002. A Novel Oral Neutrophil Elastase Inhibitor (ONO-6818) Inhibits Human Neutrophil Elastase-induced Emphysema in Rats. Am J Respir Crit Care Med. 166(4):496-500. https://doi.org/10.1164/rccm.2103118

10. Aasen AO, Ohlsson K. 1978. Release of Granulocyte Elastase in Lethal Canine Endotoxin Shock. 359(1): https://doi.org/10.1515/bchm.1978.359.1.683

11. Powers JC, Gupton B, Harley A, Nishino N, Whitley RJ. 1977. Specificity of porcine pancreatic elastase, human leukocyte elastase and cathepsin G Inhibition with peptide chloromethyl ketones. Biochimica et Biophysica Acta (BBA) - Enzymology. 485(1):156-166. https://doi.org/10.1016/0005-2744(77)90203-0

12. IUBMB Enzyme Nomenclature. [Internet]. Available from: http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/4/21/37

13. Dubin A, Potempa J, Travis J. 1994. Structural and functional characterization of elastases from horse neutrophils. 300(2):401-406. https://doi.org/10.1042/bj3000401

14. Senior RM, Bielefeld DR, Starcher BC. 1976. Comparison of the elastolytic effects of human leukocyte elastase and porcine pancreatic elastase. Biochemical and Biophysical Research Communications. 72(4):1327-1334. https://doi.org/10.1016/s0006-291x(76)80160-x

15. Roughley PJ, Barrett AJ. 1977. The degradation of cartilage proteoglycans by tissue proteinases. Proteoglycan structure and its susceptibility to proteolysis. 167(3):629-637. https://doi.org/10.1042/bj1670629

16. Starkey PM, Barrett AJ, Burleigh MC. 1977. The degradation of articular collagen by neutrophil proteinases. Biochimica et Biophysica Acta (BBA) - Enzymology. 483(2):386-397. https://doi.org/10.1016/0005-2744(77)90066-3

17. Gadek JE, Fells GA, Wright DG, Crystal RG. 1980. Human neutrophil elastase functions as a type III collagen ?Collagenase?. Biochemical and Biophysical Research Communications. 95(4):1815-1822. https://doi.org/10.1016/s0006-291x(80)80110-0

18. Davies M, Barrett AJ, Travis J, Sanders E, Coles GA. 1978. The Degradation of Human Glomerular Basement Membrane with Purified Lysosomal Proteinases: Evidence for the Pathogenic Role of the Polymorphonuclear Leucocyte in Glomerulonephritis. 54(3):233-240. https://doi.org/10.1042/cs0540233

19. McDonald J, Kelley D. 1980. Degradation of fibronectin by human leukocyte elastase. Release of biologically active fragments.. Journal of Biological Chemistry. 255(18):8848-58.

20. Ohlsson K, Odsson I. 1974. The Neutral Proteases of Human Granulocytes. Isolation and Partial Characterization of Granulocyte Elastases. Eur J Biochem. 42(2):519-527. https://doi.org/10.1111/j.1432-1033.1974.tb03367.x

21. Liau DF, Yin NX, Ryan SF. 1993. Isolation of Human Polymorphonudear Leukocyte Elastase by Chromatography on Immobilized Benzamidine. Preparative Biochemistry. 23(4):439-447. https://doi.org/10.1080/10826069308544568

22. Taylor JC, Crawford IP. 1975. Purification and preliminary characterization of human leukocyte elastase. Archives of Biochemistry and Biophysics. 169(1):91-101. https://doi.org/10.1016/0003-9861(75)90320-3