Alpha-1 antitrypsin

Serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 1
Crystal structure of serpin A1 (red) in an inhibitory complex with pancreatic elastase (blue). Rendered from PDB 2D26.
Available structures PDB Ortholog search: PDBe, RCSB List of PDB id codes 1ATU, 1D5S, 1EZX, 1HP7, 1IZ2, 1KCT, 1OO8, 1OPH, 1PSI, 1QLP, 1QMB, 2D26, 2QUG, 3CWL, 3CWM, 3DRM, 3DRU, 3NDD, 3NDF, 3NE4, 3T1P, 7API, 8API, 9API
Identifiers
Symbols	SERPINA1 ; A1A; A1AT; AAT; PI; PI1; PRO2275; alpha1AT
External IDs	OMIM: 107400 MGI: 891971 HomoloGene: 20103 GeneCards: SERPINA1 Gene
Gene ontology Molecular function • glycoprotein binding • protease binding • serine-type endopeptidase inhibitor activity • protein binding • identical protein binding Cellular component • extracellular region • proteinaceous extracellular matrix • extracellular space • endoplasmic reticulum • Golgi apparatus • platelet alpha granule lumen • extracellular exosome Biological process • platelet degranulation • acute-phase response • blood coagulation • negative regulation of endopeptidase activity • platelet activation Sources: Amigo / QuickGO
RNA expression pattern
More reference expression data
Orthologs
Species	Human	Mouse
Entrez	5265	20700
Ensembl	ENSG00000197249	ENSMUSG00000066366
UniProt	P01009	P07758
RefSeq (mRNA)	NM_000295	NM_001252569
RefSeq (protein)	NP_000286	NP_001239498
Location (UCSC)	Chr 14: 94.38 – 94.39 Mb	Chr 12: 103.85 – 103.86 Mb
PubMed search	[1]	[2]

Alpha-1 Antitrypsin or α₁-antitrypsin (A1AT) is a protease inhibitor belonging to the serpin superfamily. It is generally known as serum trypsin inhibitor. Alpha 1-antitrypsin is also referred to as alpha-1 proteinase inhibitor (A1PI) because it inhibits a wide variety of proteases. It protects tissues from enzymes of inflammatory cells, especially neutrophil elastase, and has a reference range in blood of 1.5 - 3.5 gram/liter (in US the reference range is generally expressed as mg/dL or micromoles), but the concentration can rise manyfold upon acute inflammation. In its absence (such as in alpha 1-antitrypsin deficiency), neutrophil elastase is free to break down elastin, which contributes to the elasticity of the lungs, resulting in respiratory complications such as emphysema, or COPD (chronic obstructive pulmonary disease) in adults and cirrhosis in adults or children.

Function

A1AT is a 52-kDa serpin and, in medicine, it is considered the most prominent serpin; the terms α1-antitrypsin and protease inhibitor (P_i) are often used interchangeably.

Most serpins inactivate enzymes by binding to them covalently, requiring very high levels to perform their function. In the acute phase reaction, a further elevation is required to "limit" the damage caused by activated neutrophil granulocytes and their enzyme elastase, which breaks down the connective tissue fiber elastin.

Like all serine protease inhibitors, A1AT has a characteristic secondary structure of beta sheets and alpha helices. Mutations in these areas can lead to non-functional proteins that can polymerise and accumulate in the liver (infantile hepatic cirrhosis).

Role in disease

Main article: Alpha 1-antitrypsin deficiency

Disorders of this protein include alpha 1-antitrypsin deficiency, an autosomal codominant hereditary disorder in which a deficiency of alpha 1-antitrypsin leads to a chronic uninhibited tissue breakdown. This causes the degradation especially of lung tissue, and eventually leads to characteristic manifestations of pulmonary emphysema. Evidence has shown that cigarette smoke can lead to oxidation of methionine 358 of α₁-antitrypsin (382 in the pre-processed form containing the 24 amino acid signal peptide), a residue essential for binding elastase; this is thought to be one of the primary mechanisms by which cigarette smoking (or second-hand smoke) can lead to emphysema. Because A1AT is expressed in the liver, certain mutations in the gene encoding the protein can cause misfolding and impaired secretion, which can lead to liver cirrhosis.

An extremely rare form of P_i, termed P_iPittsburgh, functions as an antithrombin (a related serpin), due to a mutation (Met358Arg). One person with this mutation has been reported to have died of a lethal bleeding diathesis.

Liver biopsy will show abundant PAS-positive globules within periportal hepatocytes.

Nomenclature

The protein was originally named "antitrypsin" because of its ability to covalently bind and irreversibly inactivate the enzyme trypsin in vitro. Trypsin, a type of peptidase, is a digestive enzyme active in the duodenum and elsewhere.

The term alpha-1 refers to the protein's behavior on protein electrophoresis. On electrophoresis, the protein component of the blood is separated by electric current. There are several clusters, the first being albumin, the second being the alpha, the third beta and the fourth gamma (immunoglobulins). The non-albumin proteins are referred to as globulins.

The alpha region can be further divided into two sub-regions, termed "1" and "2". Alpha 1-antitrypsin is the main protein of the alpha-globulin 1 region.

Another name used is alpha-1 proteinase inhibitor (α₁-PI).

Genetics

The gene is located on the long arm of the fourteenth chromosome (14q32.1).

Over 100 different variants of α₁-antitrypsin have been described in various populations. North-Western Europeans are most at risk for carrying one of the most common mutant forms of A1AT, the Z mutation (Glu342Lys on M1A, rs28929474).

Biochemical Properties

A1AT is a single-chain glycoprotein consisting of 394 amino acids in the mature form and exhibits a number of glycoforms. The three N-linked glycosylations sites are mainly equipped with so-called diantennary N-glycans. However, one particular site shows a considerable amount of heterogeneity since tri- and even tetraantennary N-glycans can be attached to the Asparagine 107 (ExPASy amino acid nomenclature). These glycans carry different amounts of negatively charged sialic acids, this causes the heterogeneity observed on normal A1AT when analysed by isoelectric focussing. In addition, the fucosylated triantennary N-glycans were shown to have the fucose as part of a so-called Sialyl Lewis x epitope, which could confer this protein particular protein-cell recognition properties. The single cysteine residue of A1AT in position 256 (ExPASy nomenclature) is found to be covalently linked to a free single cysteine by a disulfide bridge.

Analysis

The level of A1AT in serum is most often determined by adding an antibody that binds to A1AT, then using turbidimetry to measure how much A1AT is present. Other detection methods include use of enzyme-linked-immuno-sorbent-assays and radial immunodiffusion.

Different analytical methods are used to determine A1AT phenotype. As protein electrophoresis is imprecise, A1AT phenotype is analysed by isoelectric focusing (IEF) in the pH range 4.5-5.5, where the protein migrates in a gel according to its isoelectric point or charge in a pH gradient.

Normal A1AT is termed M, as it is migrates toward the center of such an IEF gel. Other variants are less functional, and are termed A-L and N-Z, dependent on whether they run proximal or distal to the M band. The presence of deviant bands on IEF can signify the presence of alpha 1-antitrypsin deficiency. Since the number of identified mutations has exceeded the number of letters in the alphabet, subscripts have been added to most recent discoveries in this area, as in the Pittsburgh mutation described above.

As every person has two copies of the A1AT gene, a heterozygote with two different copies of the gene may have two different bands showing on electrofocusing, although heterozygote with one null mutant that abolishes expression of the gene will only show one band.

In blood test results, the IEF results are notated as in P_iMM, where P_i stands for protease inhibitor and "MM" is the banding pattern of that patient.

Alpha 1-antitrypsin levels in the blood depend on the genotype. Some mutant forms fail to fold properly and are, thus, targeted for destruction in the proteasome, whereas others have a tendency to polymerise, being retained in the endoplasmic reticulum. The serum levels of some of the common genotypes are:

• PiMM: 100% (normal)
• PiMS: 80% of normal serum level of A1AT
• PiSS: 60% of normal serum level of A1AT
• PiMZ: 60% of normal serum level of A1AT
• PiSZ: 40% of normal serum level of A1AT
• PiZZ: 10-15% (severe alpha 1-antitrypsin deficiency)

• PiZ is caused by a glutamate to lysine mutation at position 342 (366 in pre-processed form)
• PiS is caused by a glutamate to valine mutation at position 264 (288 in pre-processed form)

Other rarer forms have been described; in all there are over 80 variants.

Therapeutic use

Alpha-1 antitrypsin

Systematic (IUPAC) name
Alpha-1-proteinase inhibitor
Clinical data
Routes of administration	intravenous
Identifiers
CAS Number	9041-92-3 Y
ATC code	B02AB02 (WHO)
DrugBank	DB00058 Y
Chemical data
Formula	C2001H3130N514O601S10
Molecular mass	44324.5 g/mol
(verify)

Recombinant alpha 1-antitrypsin is not yet commercially available, but is under investigation as a therapy for alpha 1-antitrypsin deficiency.

Therapeutic concentrates are prepared from the blood plasma of blood donors. The US FDA has approved the use of three alpha 1-antitrypsin products derived from a human plasma: Prolastin, Zemaira, and Aralast. These products for intravenous augmentation A1AT therapy can cost up to $100,000 per year per patient. They are administered intravenously at a dose of 60 mg/kg once a week.

A recent study analyzed and compared the three FDA-approved products in terms of their primary structure and glycosylation. All three products showed minor differences compared to the normal human plasma A1AT, and are introduced during the specific purifications procedures. However, these detected differences are not believed to have any negative implications to the patients.

Aerosolized-augmented A1AT therapy is under study. This involves inhaling purified human A1AT into the lungs and trapping the A1AT into the lower respiratory tract. This method proves more successful than intravenous-augmented A1AT therapy because intravenous use of A1AT results in only 10%-15% of the A1AT reaching the lower respiratory tract, whereas 25%-45% of A1AT can reach the lower respiratory tract through inhalation. However, inhaled A1AT may not reach the elastin fibers in the lung where elastase injury actually occurs. Further study is currently underway.

History

The possibility of allelic variants of A1AT leading to disease was first investigated by Axelsson and Laurell in 1965.

See also

• Trypsin inhibitor

External links

• The MEROPS online database for peptidases and their inhibitors: I04.001
• Proteopedia: Alpha-1-antitrypsin
• Alpha 1-antitrypsin at Lab Tests Online