Pancreatic elastase

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Pancreatic elastase
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ExPASy
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KEGGKEGG entry
MetaCycmetabolic pathway
PRIAMprofile
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Pancreatic elastase II
Identifiers
ExPASy
NiceZyme view
KEGGKEGG entry
MetaCycmetabolic pathway
PRIAMprofile
PDB structuresRCSB PDB PDBe PDBsum
Gene OntologyAmiGO / QuickGO
Search
PMCarticles
PubMedarticles
NCBIproteins
Pancreatic endopeptidase E
Identifiers
ExPASy
NiceZyme view
KEGGKEGG entry
MetaCycmetabolic pathway
PRIAMprofile
PDB structuresRCSB PDB PDBe PDBsum
Search
PMCarticles
PubMedarticles
NCBIproteins

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

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

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

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-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

Mutations of the CELA1 gene were suspected to be associated with

KRT6C gene has been linked to some cases of diffuse NEPPK.[7]

A possible

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

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

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

Glycosylation at Asn79 and Asn233.[9]

Gene

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

Diffuse nonepidermolytic palmoplantar keratoderma.[3]

Reactions

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

Human pancreatic elastase 1 (E1) is not degraded in intestinal transit, so that its concentration in feces reflects exocrine pancreatic function. In 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:

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

External links

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