Pancreatic elastase

Identifiers
EC no.| 3.4.21.36
CAS no.| 848900-32-3
Databases
IntEnz| IntEnz view
BRENDA| BRENDA entry
ExPASy| NiceZyme view
KEGG| KEGG entry
MetaCyc| metabolic pathway
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PDB structures| RCSB PDB PDBe PDBsum
Gene Ontology| AmiGO  / QuickGO
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PubMed| articles
NCBI| proteins
Pancreatic elastase II

Identifiers
EC no.| 3.4.21.71
CAS no.| 75603-19-9
Databases
IntEnz| IntEnz view
BRENDA| BRENDA entry
ExPASy| NiceZyme view
KEGG| KEGG entry
MetaCyc| metabolic pathway
PRIAM| profile
PDB structures| RCSB PDB PDBe PDBsum
Gene Ontology| AmiGO  / QuickGO
| Search

PMC| articles
PubMed| articles
NCBI| proteins
Pancreatic endopeptidase E

Identifiers
EC no.| 3.4.21.70
CAS no.| 68073-27-8
Databases
IntEnz| IntEnz view
BRENDA| BRENDA entry
ExPASy| NiceZyme view
KEGG| KEGG entry
MetaCyc| metabolic pathway
PRIAM| profile
PDB structures| RCSB PDB PDBe PDBsum
| Search

PMC| articles
PubMed| articles
NCBI| proteins

Pancreatic elastase is a form of elastase that is produced in the acinar cells of the pancreas, initially produced as an inactive zymogen and later activated in the duodenum by trypsin. Elastases form a subfamily of serine proteases, characterized by a distinctive structure consisting of two beta barrel domains converging at the active site that hydrolyze amides and esters amongst many proteins in addition to elastin, a type of connective tissue that holds organs together. Pancreatic elastase 1 is a serine endopeptidase, a specific type of protease that has the amino acid serine at its active site. Although the recommended name is pancreatic elastase, it can also be referred to as elastase-1, pancreatopeptidase, PE, or serine elastase.

The first isozyme, pancreatic elastase 1, was initially thought to be expressed in the pancreas. However it was later discovered that it was the only chymotrypsin-like elastase that was not expressed in the pancreas. In fact, pancreatic elastase is expressed in basal layers of epidermis (at protein level). Hence pancreatic elastase 1 has been renamed elastase 1 (ELA1) or chymotrypsin-like elastase family, member 1 (CELA1).[1] For a period of time, it was thought that ELA1 / CELA1 was not transcribed into a protein.[2] However it was later discovered that it was expressed in skin keratinocytes.[3]

Clinical literature that describes human elastase 1 activity in the pancreas or fecal material is actually referring to chymotrypsin-like elastase family, member 3B (CELA3B).[1]

## Structure[edit]

Pancreatic elastase is a compact globular protein with a hydrophobic core. This enzyme is formed by three subunits. Each subunit binds one calcium ion (cofactor). There are three important metal-binding sites in amino acids 77, 82, 87.[4] The catalytic triad , located in the active site is formed by three hydrogen-bonded amino acid residues (H71, D119, S214), and plays an essential role in the cleaving ability of all proteases. It is composed of a single peptide chain of 240 amino acids and contains 4 disulfide bridges. It has a high degree of sequence identity with pancreatic elastases that correspond to other species, such as the rat's, with whom it shares 86% of its sequence.[5] Its enzymatic activity is a result of the specific three-dimensional conformation which its single polypeptide chain adopts, and therefore, activity is lost by denaturation and/or conformational changes.[citation needed]

## Inhibitors[edit]

Elafin, the skin-derived elastase inhibitor, has been shown to be a potent and specific inhibitor of both the porcine homolog of ELA1 and human leukocyte elastase in vitro. Elafin is expressed by epidermal keratinocytes under hyperproliferative conditions such as psoriasis and wound healing. It has also been reported to be present in many other adult epithelia that are exposed to environmental stimuli: tongue, plate, lingual tonsils, gingiva, pharynx, epiglottis, vocal fold, esophagus, uterine cervix, vagina, and hair follicles. In all these tissues, the presence of inflammatory cells is physiologic and elafin expression is believed to protect against leukocyte proteases, thereby helping to maintain epithelial integrity.[citation needed]

Elafin on the contrary has never been found in the basal layer in any type of epithelial tissue. Indeed, elafin is virtually absent in normal human epidermis. The other known elastase inhibitor, SLPI, however, has been reported to be expressed in the basal keratinocytes suggesting that this may be the major elastase inhibitor in normal epidermis.[citation needed]

Alpha 1-antitrypsin and alpha-2-macroglobulin are human serum protease inhibitors that completely inhibit the general proteolytic activity of pancreatic elastase 1 and 2. It has been observed that a protease must be active in order to bind to these two inhibitors. Studies proved that the activity of elastase 2 was enhanced in 25-250 mM NaCl. The activity of elastase 2 in NaCl approached approximately twice the activity without NaCl. Elastase 1 is slightly inhibited above 150 mM NaCl[6]

## Clinical significance[edit]

Mutations of the CELA1 gene were suspected to be associated with diffuse nonepidermolytic palmoplantar keratoderma (diffuse NEPPK).[3] However the suspected sequence variant was fully functional and did not strongly associate with the disease. More recently, a specific mutation in the KRT6C gene has been linked to some cases of diffuse NEPPK.[7]

A possible polymorphism of the CELA1 gene coding this protein was found. On a secondary structure level, this polymorphism manifests itself in an excision of a short sequence of CELA1. The disappeared sequence carries the key amino acid residues Val-227 and Thr-239, which contribute to the substrate specificity of elastase I (highlighted in Figure 3), as well as five of the eight amino acids involved in the primary contact of the elafin(inhibitor)/elastase complex formation. These observations imply that the sequence variant might modify the substrate specificity of the enzyme and abolish the inhibitor binding capability. Though there were no obvious pathogenic epidermal abnormalities associated with the truncated ELA1 variant, it is possible that carriers of the polymorphism may be at greater risk of developing the common skin diseases such as psoriasis and eczema (genetic and histologic studies will be required to investigate the role of ELA1 in these common epidermal disorders.).[3]

## Biosynthesis[edit]

Pancreatic elastase is formed by activation of proelastase from mammalian pancreas by trypsin. After processing to proelastase, it is stored in the zymogen granules and then activated to elastase in the duodenum by the tryptic cleavage of a peptide bond in the inactive form of the precursor molecule.[8] This process results in the removal of an activation peptide from the N-terminal, that enables the enzyme to adopt its native conformation.[citation needed]

## Isozymes[edit]

Humans have five chymotrypsin-like elastase genes which encode the structurally similar proteins:

Family  | Gene symbol  | Protein name  | EC number

|
|
|

Approved  | Previous  | Approved  | Previous
chymotrypsin-
like  | CELA1 | ELA1  | chymotrypsin-like elastase family, member 1  | elastase 1, pancreatic  | EC 3.4.21.36
CELA2A | ELA2A  | chymotrypsin-like elastase family, member 2A  | elastase 2A, pancreatic  | EC 3.4.21.71
CELA2B | ELA2B  | chymotrypsin-like elastase family, member 2B  | elastase 2B, pancreatic  | EC 3.4.21.71
CELA3A | ELA3A  | chymotrypsin-like elastase family, member 3A  | elastase 3A, pancreatic  | EC 3.4.21.70
CELA3B | ELA3B  | chymotrypsin-like elastase family, member 3B  | elastase 3B, pancreatic  | EC 3.4.21.70

## Post-translational modifications[edit]

Glycosylation at Asn79 and Asn233.[9]

## Gene[edit]

The gene that codes for pancreatic elastase 1 is CELA1 (synonym: ELA1) Pancreatic elastase 1 is encoded by a single genetic locus on chromosome 12. Studies of human pancreatic elastase 1 have shown that this serine protease maps to the chromosomal region 12q13[10] and it is close to a locus for an autosomal dominant skin disease, Diffuse nonepidermolytic palmoplantar keratoderma.[3]

## Reactions[edit]

Reaction catalysed by pancreatic elastase 1. This image represents the hydrolysis of the succinyl-Ala-Ala-Ala-p-nitroanalide. The addition of one water molecule provokes the hydrolysis of the molecule and the release of p-nitroaniline.

The hydrolysis that elastases bring about occur in several steps, starting with the formation of a complex between elastase and its substrate, with the carbonyl carbon positioned near the nucleophilic serine, followed by a nucleophillic attack that forms an acyl-enzyme intermediate (a pair of electrons from the double bond of the carbonyl oxygen moves to the oxygen) while the first product is released. The intermediate is then hydrolyzed in a deacylation step, regenerating the active enzyme and resulting in the release of the second product ( the electron-deficient carbonyl carbon re-forms the double bond with the oxygen and the C-terminus of the peptide is released. It preferentially cleaves peptide bonds at the carbonyl end of amino acid residues with small hydrophobic side chains such as glycine, valine, leucine, isoleucine and alanine. The wide specificity of elastases for non-aromatic uncharged side chains can explain its ability to break down native elastin.[11]

## Use in diagnostic tests[edit]

Human pancreatic elastase 1 (E1) remains undegraded during intestinal transit. Therefore, its concentration in feces reflects exocrine pancreatic function. During an inflammation of the pancreas, E1 is released into the bloodstream. Thus the quantification of pancreatic elastase 1 in serum allows diagnosis or exclusion of acute pancreatitis.[12]

Main indications:

* Diagnosis/exclusion of exocrine pancreatic insufficiency caused by e.g. Chronic Pancreatitis, Cystic Fibrosis, Diabetes Mellitus, Cholelithiasis (Gallstones), "Failure to Thrive", Pancreatic Cancer, Papillary Stenosis
* Follow-up monitoring of patients with mild or moderate pancreatic insufficiency
* Diagnosis/exclusion of pancreatic involvement in association with gastrointestinal symptoms, abdominal pain or osteoporosis, for example.[13]

Method of detection:

* Sandwich ELISA with two monoclonal antibodies highly specific for human pancreatic elastase 1
* The ELISA kit is based on a microtiter plate (96 well format) with 12 breakable single strips x 8 wells suitable for up to 42 samples in duplicate

Reference concentration to interpret Pancreatic Elastase results: For adults and children after the first month of life

* Values > 200 μg elastase/g stool indicate normal exocrine pancreatic function
* Values of 100-200 μg elastase/g stool suggest mild to moderate pancreatic insuffiency
* Values < 100 μg elastase/g stool indicate exocrine pancreatic insufficiency.[14]

## References[edit]

1. ^ a b EntrezGene 1990
2. ^ Rose SD, MacDonald RJ (June 1997). "Evolutionary silencing of the human elastase I gene (ELA1)". Hum. Mol. Genet. 6 (6): 897–903. doi:10.1093/hmg/6.6.897. PMID 9175736.
3. ^ a b c d Talas U, Dunlop J, Khalaf S, Leigh IM, Kelsell DP (January 2000). "Human elastase 1: evidence for expression in the skin and the identification of a frequent frameshift polymorphism". J. Invest. Dermatol. 114 (1): 165–70. doi:10.1046/j.1523-1747.2000.00825.x. PMID 10620133.
4. ^ Universal protein resource accession number Q9UNI1 at UniProt.
5. ^ "Elastase". Worthington Enzyme Manual.
6. ^ Largman C, Brodrick JW, Geokas MC (1976). "Purification and characterization of two human pancreatic elastases". Biochemistry. 15 (11): 2491–500. doi:10.1021/bi00656a036. PMID 819031.
7. ^ Akasaka E, Nakano H, Nakano A, Toyomaki Y, Takiyoshi N, Rokunohe D, Nishikawa Y, Korekawa A, Matsuzaki Y, Mitsuhashi Y, Sawamura D (2011). "Diffuse and focal palmoplantar keratoderma can be caused by a keratin 6c mutation". Br. J. Dermatol. 165 (6): 1290–2. doi:10.1111/j.1365-2133.2011.10552.x. PMID 21801157. S2CID 36039184.
8. ^ Gertler A, Birk Y (1970). "Isolation and characterization of porcine proelastase". Eur. J. Biochem. 12 (1): 170–6. doi:10.1111/j.1432-1033.1970.tb00835.x. PMID 5461547.
9. ^ "CELA1 Gene". GeneCards.
10. ^ Davies RL, Yoon SJ, Weissenbach J, Ward D, Krauter K, Kucherlapati R (October 1995). "Physical mapping of the human ELA1 gene between D12S361 and D12S347 on chromosome 12q13". Genomics. 29 (3): 766–8. doi:10.1006/geno.1995.9939. PMID 8575772.
11. ^ Shotton DM (1970). "[7] Elastase". Elastase. Methods in Enzymology. Vol. 19\. pp. 113–140. doi:10.1016/0076-6879(70)19009-4. ISBN 9780121818814.
12. ^ Stein J, Schoonbroodt D, Jung M, Lembcke B, Caspary WF (1996). "Mesure de l'élastase fécale par immunoréactivité: une nouvelle approche indirecte de la fonction pancréatique" [Measurement of fecal elastase 1 by immunoreactivity: A new indirect test of the pancreatic function]. Gastroentérologie Clinique et Biologique (in French). 20 (5): 424–9. PMID 8761139.
13. ^ Gonzales AC, Vieira SM, Maurer RL, Silva FA, Silveira TR (2011). "Use of monoclonal faecal elastase-1 concentration for pancreatic status assessment in cystic fibrosis patients". J Pediatr (Rio J). 87 (2): 157–62. doi:10.2223/JPED.2075. PMID 21503378.
14. ^ Löser C, Möllgaard A, Fölsch UR (October 1996). "Faecal elastase 1: a novel, highly sensitive, and specific tubeless pancreatic function test". Gut. 39 (4): 580–6. doi:10.1136/gut.39.4.580. PMC 1383273. PMID 8944569.

## External links[edit]

* Pancreatic+Elastase at the U.S. National Library of Medicine Medical Subject Headings (MeSH)
* Overview of all the structural information available in the PDB for UniProt: P00772 (Pancreatic elastase) at the PDBe-KB.

This article incorporates text from the United States National Library of Medicine, which is in the public domain.

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Endopeptidases: serine proteases/serine endopeptidases (EC 3.4.21)

Digestive enzymes|

* Enteropeptidase
* Trypsin
* Chymotrypsin
* Elastase
* Neutrophil
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Enzymes

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