ERAP2

Expression ERAP2 expression is regulated by interferon gamma signalling. While ERAP2 and homologous enzyme ERAP1 are both expressed in immune cells, the expression of the enzymes is independently regulated in other tissues without significant correlation of expression levels. However, coordinated expression patterns have also been observed, in which ERAP2 downregulation is counterbalanced by an increase in ERAP1 expression. Overexpression of ERAP2 in various cancer types, including melanomas and different adenocarcinomas, has been suggested to modulate the presentation of cancer antigens on MHC-I, which may affect cancer invasion by immune cells. ERAP2 is not expressed in mice making it more difficult to study. Antigen presentation Unlike ERAP1, ERAP2 can trim efficiently peptides that already have optimal length for MHC class I presentation. Thus ERAP2 has been shown to shorten peptides of 9 or fewer amino acids, thereby destroying antigenic peptides in some cases. Beyond N-terminal trimming, ERAP2 has also been reported to influence the immunopeptidome through internal sequence preferences that shape peptide motifs presented by specific MHC class I alleles. In the context of HLA-A29, ERAP2 expression promotes the generation of a distinct HLA-A29-specific peptide submotif. This peptide motif is detectable in putative autoantigens implicated in HLA-A29-associated birdshot uveitis, a disease strongly associated with genetic variation in ERAP2 and ERAP1. Analyses of the HLA-A29 immunopeptidome further indicate that ERAP2-dependent peptide selection reflects internal sequence specificity beyond simple N-terminal trimming. ERAP2 displays a preference for peptide substrates that carry N-terminal basic residues (arginine, lysine). Heterodimer formation improves peptide-trimming efficiency, resulting in an expanded antigenic repertoire and a more diverse immune response. The ERAP1-ERAP2 complex can trim free peptides in the ER and may also be able to trim MHC I-bound precursor peptides, according to some authors. Individuals homozygous for ERAP2 haplotype B lack ERAP2 protein expression and have significantly lower MHC class I levels on the surface of B cells. This may result in an altered presentation of antigens and resulting immune responses. Other functions ERAP2 can modulate the renin-angiotensin system (RAS) in blood pressure homeostasis through angiotensin cleavage. In concert with ERAP1, ERAP2 counteracts angiotensin II activity, inducing vasodilation and hypertension reduction. Blood pressure modulation by ERAP2 is supported by the association of ERAP2 with blood pressure progression and hypertension incidence. Its link to pre-eclampsia in multiple populations shows further support for the role of ERAP2 in blood pressure homeostasis. == Genetics / clinical significance ==

Gene / location ERAP2 gene is located on human chromosome 5 in between the ERAP1 and LNPEP genes encoding the other two family members of the M1 aminopeptidases. It has 41,438 base pair length and consists of 19 exons. SNPs and disease association The ERAP2 gene is highly polymorphic and contains many common single nucleotide variants (SNVs) that are in strong linkage disequilibrium and are maintained at intermediate frequencies through balancing selection. There are two common SNVs in ERAP2 that facilitate the alternative splicing of three haplotypes by altering splice motifs. Under steady state conditions, the truncated mRNA is destroyed by nonsense mediated decay (NMD), but during influenza infections it is translated to two truncated isoforms. Accordingly, 25% of the general population (haplotype B homozygotes) are deficient in full-length ERAP2 protein. The G allele of SNV rs17486481 activates a cryptic splice site upstream of exon 12 that also introduces premature stop codons and makes the transcript likely vulnerable to NMD (termed haplotype CDifferent ERAP2 protein haplotypes (or allotypes) have been detected among the major continental populations based on common missense variants in ERAP2. Interestingly, the alleles from SNVs that strongly predispose to autoimmune conditions (i.e., A allele of rs2248374 and other SNVs in haplotype A) display natural selection in recent human history, Klunk et al. found that individuals with the protective allele (dominant in the present European population) had a fivefold increase in ERAP2 expression in macrophages resulting in reduced replication of Y. pestis. Large-scale genomic studies have identified SNPs in ERAP2 as key host factors influencing control of latent Epstein–Barr virus (EBV) infection. These findings suggest that variation in ERAP2 may contribute to differences in host control of EBV latency and viral DNA persistence. Given the established role of EBV in B-cell biology, genetic associations between EBV activity and autoimmune diseases such as multiple sclerosis and systemic lupus erythematosus (SLE) indicate shared host pathways linking viral persistence and immune-mediated disease. == Structure / Mechanism ==

Structure ERAP2 is composed of 4 structural units (I-IV), with the HEXXHX18E zinc-binding motif and the known GAMEN aminopeptidase motif located in domain II, similarly to its closely related enzyme ERAP1. The catalytic site features a single Zn(II) ion and is coordinated by two histidine residues (H370, H374) and a glutamate residue (E393). Domains II and IV, which are linked by domain III, form a large internal cavity close to the catalytic site and exclude the external solvent, in accordance with the "closed" conformation obtained for ERAP1. Mechanism ERAP2 selects substrates by sequestering them in its internal cavity and allowing interactions to determine trimming rates, thus combining substrate permissiveness with sequence bias. A crystal structure of ERAP2 with a peptide product located in this cavity has revealed lack of deep specificity pockets and lack of a cavity that interacts with the peptide C- terminus, which justify the limited selectivity of this enzyme and the differences in length selection compared to ERAP1 (ERAP2 can effectively trim 8-mer peptides, while it is less active with longer substrates In terms of N-terminal residue specificity, ERAP2 prefers basic amino acids, such as arginine. == Therapeutic approaches and pharmacology ==

Therapeutic approaches for ERAP2 regulation rely mostly on the development of small molecule inhibitors. The most explored classes of inhibitors for ERAP2 are the allosteric site ones. (HTS) against ERAP2, from an in-house library of 1920 compounds. Compound 3 was amongst those selected for their potency (ERAP2, IC50 = 22 nM) and selectivity. Docking studies revealed that the carboxylic acid is predicted to coordinate the catalytic zinc ion within ERAP2. Several analogues were designed and synthesized. ERAP2 allosteric site inhibitors • Sulfonamides Sulfonamide compound 4 was identified as a potential allosteric inhibitor against ERAP2 in 2022 by Arya et al.. This compound targets ERAP2 through an uncompetitive manner (IC50 = 44 μM) by inhibiting the hydrolysis of peptide substrates. At the same time it acts as a competitive inhibitor against ERAP1 (IC50 = 73 μM). == References ==