{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T13:16:53Z","timestamp":1762521413620,"version":"build-2065373602"},"reference-count":52,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2011,5,24]],"date-time":"2011-05-24T00:00:00Z","timestamp":1306195200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Marine Drugs"],"abstract":"<jats:p>Arylalkylamine N-acetyltransferase (AANAT) catalyzes the transfer of an acetyl group from acetyl coenzyme A (AcCoA) to arylalkylamines, including indolethylamines and phenylethylamines. Multiple aanats are present in teleost fish as a result of whole genome and gene duplications. Fish aanat1a and aanat2 paralogs display different patterns of tissue expression and encode proteins with different substrate preference: AANAT1a is expressed in the retina, and acetylates both indolethylamines and phenylethylamines; while AANAT2 is expressed in the pineal gland, and preferentially acetylates indolethylamines. The two enzymes are therefore thought to serve different roles. Here, the molecular changes that led to their specialization were studied by investigating the structure-function relationships of AANATs in the gilthead seabream (sb, Sperus aurata). Acetylation activity of reciprocal mutated enzymes pointed to specific residues that contribute to substrate specificity of the enzymes. Inhibition tests followed by complementary analyses of the predicted three-dimensional models of the enzymes, suggested that both phenylethylamines and indolethylamines bind to the catalytic pocket of both enzymes. These results suggest that substrate selectivity of AANAT1a and AANAT2 is determined by the positioning of the substrate within the catalytic pocket, and its accessibility to catalysis. This illustrates the evolutionary process by which enzymes encoded by duplicated genes acquire different activities and play different biological roles.<\/jats:p>","DOI":"10.3390\/md9050906","type":"journal-article","created":{"date-parts":[[2011,5,25]],"date-time":"2011-05-25T05:09:13Z","timestamp":1306300153000},"page":"906-921","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Molecular Evolution of Multiple Arylalkylamine N-Acetyltransferase (AANAT) in Fish"],"prefix":"10.3390","volume":"9","author":[{"given":"Bina","family":"Zilberman-Peled","sequence":"first","affiliation":[{"name":"Department of Neurobiology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, 69978, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sharron","family":"Bransburg-Zabary","sequence":"additional","affiliation":[{"name":"Bioinformatics Unit, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, 69978, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David C.","family":"Klein","sequence":"additional","affiliation":[{"name":"Section on Neuroendocrinology, Office of the Scientific Directory, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoav","family":"Gothilf","sequence":"additional","affiliation":[{"name":"Department of Neurobiology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, 69978, Israel"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2011,5,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1016\/S0968-0004(97)01034-7","article-title":"GCN5-related histone N-acetyltransferases belong to a diverse superfamily that includes the yeast SPT10 protein","volume":"22","author":"Neuwald","year":"1997","journal-title":"Trends Biochem Sci"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1146\/annurev.biophys.29.1.81","article-title":"GCN5-related N-acetyltransferases: A structural overview","volume":"29","author":"Dyda","year":"2000","journal-title":"Annu Rev Biophys Biomol Struct"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.pep.2004.07.004","article-title":"Purification of the recombinant human serotonin N-acetyltransferase (EC 2.3.1.87): Further characterization of and comparison with AANAT from other species","volume":"38","author":"Ferry","year":"2004","journal-title":"Protein Expr Purif"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"30321","DOI":"10.1074\/jbc.273.46.30321","article-title":"Indoleamine analogs as probes of the substrate selectivity and catalytic mechanism of serotonin N-acetyltransferase","volume":"273","author":"Khalil","year":"1998","journal-title":"J Biol Chem"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1681","DOI":"10.1126\/science.270.5242.1681","article-title":"Pineal serotonin N-acetyltransferase: Expression cloning and molecular analysis","volume":"270","author":"Coon","year":"1995","journal-title":"Science"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"10913","DOI":"10.1016\/S0021-9258(18)90600-9","article-title":"Arylamine N-acetyltransferase and arylalkylamine N-acetyltransferase in the mammalian pineal gland","volume":"259","author":"Voisin","year":"1984","journal-title":"J Biol Chem"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"8794","DOI":"10.1074\/jbc.275.12.8794","article-title":"Substrate specificity and inhibition studies of human serotonin N-acetyltransferase","volume":"275","author":"Ferry","year":"2000","journal-title":"J Biol Chem"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.mce.2006.03.039","article-title":"Evolution of arylalkylamine N-acetyltransferase: Emergence and divergence","volume":"252","author":"Coon","year":"2006","journal-title":"Mol Cell Endocrinol"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"292","DOI":"10.1016\/j.tig.2004.05.007","article-title":"Evolution of cell-cell signaling in animals: Did late horizontal gene transfer from bacteria have a role?","volume":"20","author":"Iyer","year":"2004","journal-title":"Trends Genet"},{"key":"ref_10","first-page":"307","article-title":"The melatonin rhythm-generating enzyme: Molecular regulation of serotonin N-acetyltransferase in the pineal gland","volume":"52","author":"Klein","year":"1997","journal-title":"Recent Prog Horm Res"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1007\/s00441-002-0579-y","article-title":"Control of melatonin synthesis in the mammalian pineal gland: The critical role of serotonin acetylation","volume":"309","author":"Ganguly","year":"2002","journal-title":"Cell Tissue Res"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3045","DOI":"10.1074\/jbc.273.5.3045","article-title":"Kinetic analysis of the catalytic mechanism of serotonin N-acetyltransferase (EC 2.3.1.87)","volume":"273","author":"Gastel","year":"1998","journal-title":"J Biol Chem"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"18118","DOI":"10.1074\/jbc.M200595200","article-title":"Investigation of the roles of catalytic residues in serotonin N-acetyltransferase","volume":"277","author":"Scheibner","year":"2002","journal-title":"J Biol Chem"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/S1097-2765(00)80171-9","article-title":"Melatonin biosynthesis: The structure of serotonin N-acetyltransferase at 2.5 A resolution suggests a catalytic mechanism","volume":"3","author":"Hickman","year":"1999","journal-title":"Mol Cell"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1016\/S0092-8674(00)80745-X","article-title":"The structural basis of ordered substrate binding by serotonin N-acetyltransferase: Enzyme complex at 1.8 A resolution with a bisubstrate analog","volume":"97","author":"Hickman","year":"1999","journal-title":"Cell"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1016\/S0092-8674(01)00316-6","article-title":"Crystal structure of the 14-3-3zeta:serotonin N-acetyltransferase complex. A role for scaffolding in enzyme regulation","volume":"105","author":"Obsil","year":"2001","journal-title":"Cell"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1006\/jmbi.2001.5371","article-title":"X-ray crystallographic studies of serotonin N-acetyltransferase catalysis and inhibition","volume":"317","author":"Wolf","year":"2002","journal-title":"J Mol Biol"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"9076","DOI":"10.1074\/jbc.274.13.9076","article-title":"Two arylalkylamine N-acetyltransferase genes mediate melatonin synthesis in fish","volume":"274","author":"Coon","year":"1999","journal-title":"J Biol Chem"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4895","DOI":"10.1210\/endo.140.10.6975","article-title":"Zebrafish serotonin N-acetyltransferase-2: Marker for development of pineal photoreceptors and circadian clock function","volume":"140","author":"Gothilf","year":"1999","journal-title":"Endocrinology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3473","DOI":"10.1097\/00001756-200011090-00015","article-title":"Photic regulation of arylalkylamine N-acetyltransferase 1 mRNA in trout retina","volume":"11","author":"Mizusawa","year":"2000","journal-title":"Neuroreport"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1046\/j.1365-2826.2003.00993.x","article-title":"Genetic, temporal and developmental differences between melatonin rhythm generating systems in the teleost fish pineal organ and retina","volume":"15","author":"Falcon","year":"2003","journal-title":"J Neuroendocrinol"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.ygcen.2004.05.007","article-title":"Duality of serotonin-N-acetyltransferase in the gilthead seabream (Sparus aurata): Molecular cloning and characterization of recombinant enzymes","volume":"138","author":"Benhar","year":"2004","journal-title":"Gen Comp Endocrinol"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1111\/j.1749-6632.2009.04435.x","article-title":"Structural and functional evolution of the pineal melatonin system in vertebrates","volume":"1163","author":"Falcon","year":"2009","journal-title":"Ann NY Acad Sci"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/S0168-9525(99)01934-4","article-title":"Vertebrate evolution: Recent perspectives from fish","volume":"16","author":"Aparicio","year":"2000","journal-title":"Trends Genet"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"280","DOI":"10.1038\/sj.hdy.6800635","article-title":"Genome evolution and biodiversity in teleost fish","volume":"94","author":"Volff","year":"2005","journal-title":"Heredity"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1146\/annurev.cellbio.18.020402.140619","article-title":"Gene co-option in physiological and morphological evolution","volume":"18","author":"True","year":"2002","journal-title":"Annu Rev Cell Dev Biol"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1016\/j.ygcen.2009.04.026","article-title":"Current knowledge on the melatonin system in teleost fish","volume":"165","author":"Falcon","year":"2010","journal-title":"Gen Comp Endocrinol"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1097\/00001756-200002070-00006","article-title":"Melatonin synthesis: Arylalkylamine N-acetyltransferases in trout retina and pineal organ are different","volume":"11","author":"Benyassi","year":"2000","journal-title":"Neuroreport"},{"key":"ref_29","first-page":"220","article-title":"A possible new role for fish retinal serotonin-N-acetyltransferase-1 (AANAT1): Dopamine metabolism","volume":"1073\u20131074","author":"Ron","year":"2006","journal-title":"Brain Res"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1177\/0748730404267340","article-title":"The 2004 Aschoff\/Pittendrigh lecture: Theory of the origin of the pineal gland\u2014A tale of conflict and resolution","volume":"19","author":"Klein","year":"2004","journal-title":"J Biol Rhythms"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1080\/07420520500545839","article-title":"Evolution of the vertebrate pineal gland: The AANAT hypothesis","volume":"23","author":"Klein","year":"2006","journal-title":"Chronobiol Int"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4233","DOI":"10.1074\/jbc.R600036200","article-title":"Arylalkylamine N-acetyltransferase: \u201cThe Timezyme\u201d","volume":"282","author":"Klein","year":"2007","journal-title":"J Biol Chem"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1677\/jme.1.01893","article-title":"Zebrafish arylalkylamine-N-acetyltransferase genes\u2014Targets for regulation of the circadian clock","volume":"36","author":"Appelbaum","year":"2006","journal-title":"J Mol Endocrinol"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3047","DOI":"10.1111\/j.1460-9568.2007.05578.x","article-title":"Cloning and early expression pattern of two melatonin biosynthesis enzymes in the turbot (Scophthalmus maximus)","volume":"25","author":"Vuilleumier","year":"2007","journal-title":"Eur J Neurosci"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"14552","DOI":"10.1074\/jbc.M800593200","article-title":"Evidence that proline focuses movement of the floppy loop of arylalkylamine N-acetyltransferase (EC 2.3.1.87)","volume":"283","author":"Pavlicek","year":"2008","journal-title":"J Biol Chem"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1016\/j.abb.2004.09.003","article-title":"Structure and functions of the GNAT superfamily of acetyltransferases","volume":"433","author":"Vetting","year":"2005","journal-title":"Arch Biochem Biophys"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.ygcen.2008.10.007","article-title":"Cloning and expression of arylalkylamine N-acetyltranferase-2 during early development and metamorphosis in the sole Solea senegalensis","volume":"161","author":"Isorna","year":"2009","journal-title":"Gen Comp Endocrinol"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/j.ygcen.2004.01.004","article-title":"Embryonic and post-embryonic expression of arylalkylamine N-acetyltransferase and melatonin receptor genes in the eye and brain of chum salmon (Oncorhynchus keta)","volume":"136","author":"Shi","year":"2004","journal-title":"Gen Comp Endocrinol"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/S0169-328X(02)00138-9","article-title":"Elevated arylalkylamine-N-acetyltransferase (AA-NAT) gene expression in medial habenular and suprachiasmatic nuclei of hibernating ground squirrels","volume":"102","author":"Yu","year":"2002","journal-title":"Brain Res Mol Brain Res"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1178","DOI":"10.3109\/07420528.2010.496911","article-title":"Melatonin-synthesizing enzymes in pineal, retina, liver, and gut of the goldfish (Carassius): mRNA expression pattern and regulation of daily rhythms by lighting conditions","volume":"27","author":"Velarde","year":"2010","journal-title":"Chronobiol Int"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/S0076-6879(83)91010-8","article-title":"Reassessment of Ellman\u2019s reagent","volume":"91","author":"Riddles","year":"1983","journal-title":"Meth Enzymol"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"87","DOI":"10.32725\/jab.2006.008","article-title":"GOSA, a simulated annealing-based program for global optimization of nonlinear problems, also reveals transyears","volume":"4","author":"Czaplicki","year":"2006","journal-title":"J Appl Biomed"},{"key":"ref_43","unstructured":"Parvez, H, Parvez, S, and Nagatsu, T (1983). Methods in Biogenic Amine Research, Elsevier\/North Holland Biomedical Press."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/0022-2836(70)90057-4","article-title":"A general method applicable to the search for similarities in the amino acid sequence of two proteins","volume":"48","author":"Needleman","year":"1970","journal-title":"J Mol Biol"},{"key":"ref_45","unstructured":"Sankoff, D, and Kruskal, JB (1983). Time Warps, String Edits and Macromolecules: The Theory and Practice of Sequence Comparison, Addison Wesley."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1002\/prot.10550","article-title":"Using multiple structure alignments, fast model building, and energetic analysis in fold recognition and homology modeling","volume":"53","author":"Petrey","year":"2003","journal-title":"Proteins"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1107\/S0021889892009944","article-title":"PROTHECK: A program to check the steriochemical quality of protein structures","volume":"26","author":"Laskowski","year":"1993","journal-title":"J Appl Cryst"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"D217","DOI":"10.1093\/nar\/gkh095","article-title":"MODBASE, a database of annotated comparative protein structure models, and associated resources","volume":"32","author":"Pieper","year":"2004","journal-title":"Nucl Acids Res"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"W368","DOI":"10.1093\/nar\/gki464","article-title":"H++: A server for estimating pKas and adding missing hydrogens to macromolecules","volume":"33","author":"Gordon","year":"2005","journal-title":"Nucleic Acids Res"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"492","DOI":"10.1016\/S0076-6879(03)74021-X","article-title":"GRASP2: Visualization, surface properties, and electrostatics of macromolecular structures and sequences","volume":"374","author":"Petrey","year":"2003","journal-title":"Meth Enzymol"},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Guigo, R, and Gusfield, D (2002). Proceedings of the 2nd Workshop on Algorithms in Bioinformatics (WABI), Springer. Volume 2452.","DOI":"10.1007\/3-540-45784-4"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1002\/prot.10397","article-title":"Taking geometry to its edge: Fast unbound rigid (and hinge-bent) docking","volume":"52","author":"Inbar","year":"2003","journal-title":"Proteins"}],"container-title":["Marine Drugs"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1660-3397\/9\/5\/906\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:56:11Z","timestamp":1760219771000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1660-3397\/9\/5\/906"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,5,24]]},"references-count":52,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2011,5]]}},"alternative-id":["md9050906"],"URL":"https:\/\/doi.org\/10.3390\/md9050906","relation":{},"ISSN":["1660-3397"],"issn-type":[{"type":"electronic","value":"1660-3397"}],"subject":[],"published":{"date-parts":[[2011,5,24]]}}}