Present in virtually all mammalian cells, its primary function in humans is the development and maintenance of the immune system. However, the full physiological role of ADA is not yet completely understood.
ADA exists in both small form (as a monomer) and large form (as a dimer-complex). In the monomer form, the enzyme is a polypeptide chain, folded into eight strands of parallel α/β barrels, which surround a central deep pocket that is the active site. In addition to the eight central β-barrels and eight peripheral α-helices, ADA also contains five additional helices: residues 19-76 fold into three helices, located between β1 and α1 folds; and two antiparallel carboxy-terminal helices are located across the amino-terminal of the β-barrel.
The ADA active site contains a zinc ion, which is located in the deepest recess of the active site and coordinated by five atoms from His15, His17, His214, Asp295, and the substrate. Zinc is the only cofactor necessary for activity.
The substrate, adenosine, is stabilized and bound to the active site by nine hydrogen bonds. The carboxyl group of Glu217, roughly coplanar with the substrate purine ring, is in position to form a hydrogen bond with N1 of the substrate. The carboxyl group of Asp296, also coplanar with the substrate purine ring, forms hydrogen bond with N7 of the substrate. The NH group of Gly184 is in position to form a hydrogen bond with N3 of the substrate. Asp296 forms bonds both with the Zn2+ ion as well as with 6-OH of the substrate. His238 also hydrogen bonds to substrate 6-OH. The 3'-OH of the substrate ribose forms a hydrogen bond with Asp19, while the 5'-OH forms a hydrogen bond with His17. Two further hydrogen bonds are formed to water molecules, at the opening of the active site, by the 2'-OH and 3'-OH of the substrate.
Due to the recessing of the active site inside the enzyme, the substrate, once bound, is almost completely sequestered from solvent. The surface exposure of the substrate to solvent when bound is 0.5% the surface exposure of the substrate in the free state.
Two proposed mechanisms exist for ADA-catalyzed deamination: 1) stereospecific addition-elimination via tetrahedral intermediate or 2) an SN2 reaction. By either mechanism, Zn2+ as a strong electrophile activates a water molecule, which is deprotonated by the basic Asp295 to form the attacking hydroxide. His238 orients the water molecule and stabilizes the charge of the attacking hydroxide. Glu217 is protonated to donate a proton to N1 of the substrate.
The reaction is stereospecific due to the location of the zinc, Asp295, and His238 residues, which all face the B-side of the purine ring of the substrate.
Competitive inhibition has been observed for ADA, where the product inosine acts at the competitive inhibitor to enzymatic activity.
ADA is considered one of the key enzymes of purine metabolism. The enzyme has been found in bacteria, plants, invertebrates, vertebrates, and mammals, with high conservation of amino acid sequence. The high degree of amino acid sequence conservation suggests the crucial nature of ADA in the purine salvage pathway.
Primarily, ADA in humans is involved in the development and maintenance of the immune system. However, ADA association has also been observed with epithelial cell differentiation, neurotransmission, and gestation maintenance. It has also been proposed that ADA, in addition to adenosine breakdown, stimulates release of excitatory amino acids and is necessary to the coupling of A1 adenosine receptors and heterotrimeric G proteins.
Some mutations in the gene for adenosine deaminase cause it not to be expressed. The resulting deficiency is one cause of severe combined immunodeficiency (SCID). Deficient levels of ADA have also been associated with pulmonary inflammation, thymic cell death, and defective T-cell receptor signaling.
Conversely, mutations causing this enzyme to be overexpressed are one cause of hemolytic anemia .
ADA1 is found in most body cells, particularly lymphocytes and macrophages, where it is present not only in the cytosol and nucleus but also as the ecto- form on the cell membrane attached to dipeptidyl peptidase-4 (aka, CD26). ADA1 is involved mostly in intracellular activity, and exists both in small form (monomer) and large form (dimer). The interconversion of small to large forms is regulated by a 'conversion factor' in the lung.
ADA2 was first identified in human spleen. It was subsequently found in other tissues including the macrophage where it co-exists with ADA1. The two isoforms regulate the ratio of adenosine to deoxyadenosine potentiating the killing of parasites. ADA2 is found predominantly in the human plasma and serum, and exists solely as a homodimer.
ADA2 is the predominant form present in human blood plasma and is increased in many diseases, particularly those associated with the immune system: for example rheumatoid arthritis, psoriasis, and sarcoidosis. The plasma ADA2 isoform is also increased in most cancers. ADA2 is not ubiquitous but co-exists with ADA1 only in monocytes-macrophages.
Total plasma ADA can be measured using high performance liquid chromatography or enzymatic or colorimetric techniques. Perhaps the simplest system is the measurement of the ammonia released from adenosine when broken down to inosine. After incubation of plasma with a buffered solution of adenosine the ammonia is reacted with a Berthelot reagent to form a blue colour which is proportionate to the amount of enzyme activity. To measure ADA2, erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) is added prior to incubation so as to inhibit the enzymatic activity of ADA1. It is the absence of ADA1 that causes SCID.
^Daddona PE, Kelley WN (Jan 1977). "Human adenosine deaminase. Purification and subunit structure". The Journal of Biological Chemistry252 (1): 110–115. PMID13062.
^ abGlader BE, Backer K, Diamond LK (Dec 1983). "Elevated erythrocyte adenosine deaminase activity in congenital hypoplastic anemia". The New England Journal of Medicine309 (24): 1486–1490. doi:10.1056/NEJM198312153092404. PMID6646173.
^Saboury AA, Divsalar A, Jafari GA, Moosavi-Movahedi AA, Housaindokht MR, Hakimelahi GH (May 2002). "A product inhibition study on adenosine deaminase by spectroscopy and calorimetry". Journal of Biochemistry and Molecular Biology35 (3): 302–305. doi:10.5483/BMBRep.2002.35.3.302. PMID12297022.
^Moriwaki Y, Yamamoto T, Higashino K (Oct 1999). "Enzymes involved in purine metabolism--a review of histochemical localization and functional implications". Histology and Histopathology14 (4): 1321–1340. PMID10506947.
^Sanchez JJ, Monaghan G, Børsting C, Norbury G, Morling N, Gaspar HB (May 2007). "Carrier frequency of a nonsense mutation in the adenosine deaminase (ADA) gene implies a high incidence of ADA-deficient severe combined immunodeficiency (SCID) in Somalia and a single, common haplotype indicates common ancestry". Annals of Human Genetics71 (Pt 3): 336–47. doi:10.1111/j.1469-1809.2006.00338.x. PMID17181544.
da Cunha JG (1992). "[Adenosine deaminase. A pluridisciplinary enzyme]". Acta Médica Portuguesa4 (6): 315–23. PMID1807098.
Franco R, Casadó V, Ciruela F, Saura C, Mallol J, Canela EI et al. (Jul 1997). "Cell surface adenosine deaminase: much more than an ectoenzyme". Progress in Neurobiology52 (4): 283–94. doi:10.1016/S0301-0082(97)00013-0. PMID9247966.
Valenzuela A, Blanco J, Callebaut C, Jacotot E, Lluis C, Hovanessian AG et al. (1997). "HIV-1 envelope gp120 and viral particles block adenosine deaminase binding to human CD26". Advances in Experimental Medicine and Biology421: 185–92. doi:10.1007/978-1-4757-9613-1_24. PMID9330696.
Moriwaki Y, Yamamoto T, Higashino K (Oct 1999). "Enzymes involved in purine metabolism--a review of histochemical localization and functional implications". Histology and Histopathology14 (4): 1321–40. PMID10506947.
Hirschhorn R (1993). "Identification of two new missense mutations (R156C and S291L) in two ADA- SCID patients unusual for response to therapy with partial exchange transfusions". Human Mutation1 (2): 166–8. doi:10.1002/humu.1380010214. PMID1284479.
Berkvens TM, van Ormondt H, Gerritsen EJ, Khan PM, van der Eb AJ (Aug 1990). "Identical 3250-bp deletion between two AluI repeats in the ADA genes of unrelated ADA-SCID patients". Genomics7 (4): 486–90. doi:10.1016/0888-7543(90)90190-6. PMID1696926.
Aran JM, Colomer D, Matutes E, Vives-Corrons JL, Franco R (Aug 1991). "Presence of adenosine deaminase on the surface of mononuclear blood cells: immunochemical localization using light and electron microscopy". The Journal of Histochemistry and Cytochemistry39 (8): 1001–8. doi:10.1177/39.8.1856451. PMID1856451.
Bielat K, Tritsch GL (Apr 1989). "Ecto-enzyme activity of human erythrocyte adenosine deaminase". Molecular and Cellular Biochemistry86 (2): 135–42. doi:10.1007/BF00222613. PMID2770711.
Murray JL, Perez-Soler R, Bywaters D, Hersh EM (Jan 1986). "Decreased adenosine deaminase (ADA) and 5'nucleotidase (5NT) activity in peripheral blood T cells in Hodgkin disease". American Journal of Hematology21 (1): 57–66. doi:10.1002/ajh.2830210108. PMID3010705.
Wiginton DA, Kaplan DJ, States JC, Akeson AL, Perme CM, Bilyk IJ et al. (Dec 1986). "Complete sequence and structure of the gene for human adenosine deaminase". Biochemistry25 (25): 8234–44. doi:10.1021/bi00373a017. PMID3028473.
Akeson AL, Wiginton DA, Dusing MR, States JC, Hutton JJ (Nov 1988). "Mutant human adenosine deaminase alleles and their expression by transfection into fibroblasts". The Journal of Biological Chemistry263 (31): 16291–6. PMID3182793.
Glader BE, Backer K (Feb 1988). "Elevated red cell adenosine deaminase activity: a marker of disordered erythropoiesis in Diamond-Blackfan anaemia and other haematologic diseases". British Journal of Haematology68 (2): 165–8. doi:10.1111/j.1365-2141.1988.tb06184.x. PMID3348976.
Daddona PE, Shewach DS, Kelley WN, Argos P, Markham AF, Orkin SH (Oct 1984). "Human adenosine deaminase. cDNA and complete primary amino acid sequence". The Journal of Biological Chemistry259 (19): 12101–6. PMID6090454.
Valerio D, Duyvesteyn MG, Meera Khan P, Geurts van Kessel A, de Waard A, van der Eb AJ (Nov 1983). "Isolation of cDNA clones for human adenosine deaminase". Gene25 (2-3): 231–40. doi:10.1016/0378-1119(83)90227-5. PMID6198240.