The expression of GALT is controlled by the actions of the FOXO3 gene. The absence of this enzyme results in classic galactosemia in humans and can be fatal in the newborn period if lactose is not removed from the diet. The pathophysiology of galactosemia has not been clearly defined.[5]
GALT catalyzes the second reaction of the Leloir pathway of galactose metabolism through ping pong bi-bi kinetics with a double displacement mechanism.[6] This means that the net reaction consists of two reactants and two products (see the reaction above) and it proceeds by the following mechanism: the enzyme reacts with one substrate to generate one product and a modified enzyme, which goes on to react with the second substrate to make the second product while regenerating the original enzyme.[7] In the case of GALT, the His166 residue acts as a potent nucleophile to facilitate transfer of a nucleotide between UDP-hexoses and hexose-1-phosphates.[8]
Two-step action of galactose-1-phosphate uridylyltransferase. Image adapted from [9]
Structural studies[]
The three-dimensional structure at 180 pm resolution (x-ray crystallography) of GALT was determined by Wedekind, Frey, and Rayment, and their structural analysis found key amino acids essential for GALT function.[8] Among these are Leu4, Phe75, Asn77, Asp78, Phe79, and Val108, which are consistent with residues that have been implicated both in point mutation experiments as well as in clinical screening that play a role in human galactosemia.[8][10]
Clinical significance[]
Deficiency of GALT causes classic galactosemia. Galactosemia is an autosomal recessive inherited disorder detectable in newborns and childhood.[11] It occurs at approximately 1 in every 40,000-60,000 live-born infants. Classical galactosemia (G/G) is caused by a deficiency in GALT activity, whereas the more common clinical manifestations, Duarte (D/D) and the Duarte/Classical variant (D/G) are caused by the attenuation of GALT activity.[12] Symptoms include ovarian failure, developmental coordination disorder (difficulty speaking correctly and consistently),[13] and neurologic deficits.[12] A single mutation in any of several base pairs can lead to deficiency in GALT activity.[14] For example, a single mutation from A to G in exon 6 of the GALT gene changes Glu188 to an arginine and a mutation from A to G in exon 10 converts Asn314 to an aspartic acid.[12] These two mutations also add new restriction enzyme cut sites, which enable detection by and large-scale population screening with PCR (polymerase chain reaction).[12] Screening has mostly eliminated neonatal death by G/G galactosemia, but the disease, due to GALT’s role in the biochemical metabolism of ingested galactose (which is toxic when accumulated) to the energetically useful glucose, can certainly be fatal.[11][15] However, those afflicted with galactosemia can live relatively normal lives by avoiding milk products and anything else containing galactose (because it cannot be metabolized), but there is still the potential for problems in neurological development or other complications, even in those who avoid galactose.[16]
^Wong LJ, Frey PA (September 1974). "Galactose-1-phosphate uridylyltransferase: rate studies confirming a uridylyl-enzyme intermediate on the catalytic pathway". Biochemistry. 13 (19): 3889–94. doi:10.1021/bi00716a011. PMID4606575.
^"Archived copy". Archived from the original on 2016-03-03. Retrieved 2010-05-19.{{cite web}}: CS1 maint: archived copy as title (link)
^ abcdWedekind JE, Frey PA, Rayment I (September 1995). "Three-dimensional structure of galactose-1-phosphate uridylyltransferase from Escherichia coli at 1.8 A resolution". Biochemistry. 34 (35): 11049–61. doi:10.1021/bi00035a010. PMID7669762.
^"Archived copy". Archived from the original on 2008-12-04. Retrieved 2010-05-19.{{cite web}}: CS1 maint: archived copy as title (link)
^ abFridovich-Keil JL (December 2006). "Galactosemia: the good, the bad, and the unknown". Journal of Cellular Physiology. 209 (3): 701–5. doi:10.1002/jcp.20820. PMID17001680. S2CID32233614.
^ abcdElsas LJ, Langley S, Paulk EM, Hjelm LN, Dembure PP (1995). "A molecular approach to galactosemia". European Journal of Pediatrics. 154 (7 Suppl 2): S21-7. doi:10.1007/BF02143798. PMID7671959. S2CID11937698.
^"Archived copy". Archived from the original on 2006-02-28. Retrieved 2010-05-19.{{cite web}}: CS1 maint: archived copy as title (link)
Tyfield L, Reichardt J, Fridovich-Keil J, Croke DT, Elsas LJ, Strobl W, Kozak L, Coskun T, Novelli G, Okano Y, Zekanowski C, Shin Y, Boleda MD (1999). "Classical galactosemia and mutations at the galactose-1-phosphate uridyl transferase (GALT) gene". Human Mutation. 13 (6): 417–30. doi:10.1002/(SICI)1098-1004(1999)13:6<417::AID-HUMU1>3.0.CO;2-0. PMID10408771.
Reichardt JK, Belmont JW, Levy HL, Woo SL (March 1992). "Characterization of two missense mutations in human galactose-1-phosphate uridyltransferase: different molecular mechanisms for galactosemia". Genomics. 12 (3): 596–600. doi:10.1016/0888-7543(92)90453-Y. PMID1373122.
Leslie ND, Immerman EB, Flach JE, Florez M, Fridovich-Keil JL, Elsas LJ (October 1992). "The human galactose-1-phosphate uridyltransferase gene". Genomics. 14 (2): 474–80. doi:10.1016/S0888-7543(05)80244-7. PMID1427861.
Reichardt JK, Levy HL, Woo SL (June 1992). "Molecular characterization of two galactosemia mutations and one polymorphism: implications for structure-function analysis of human galactose-1-phosphate uridyltransferase". Biochemistry. 31 (24): 5430–3. doi:10.1021/bi00139a002. PMID1610789.
Flach JE, Reichardt JK, Elsas LJ (August 1990). "Sequence of a cDNA encoding human galactose-1-phosphate uridyl transferase". Molecular Biology & Medicine. 7 (4): 365–9. PMID2233247.
Reichardt JK, Berg P (April 1988). "Cloning and characterization of a cDNA encoding human galactose-1-phosphate uridyl transferase". Molecular Biology & Medicine. 5 (2): 107–22. PMID2840550.
Bergren WG, Donnell GN (July 1973). "A new variant of galactose-1-phosphate uridyltransferase in man: the Los Angeles variant". Annals of Human Genetics. 37 (1): 1–8. doi:10.1111/j.1469-1809.1973.tb01808.x. PMID4759900. S2CID22699183.
Shih LY, Suslak L, Rosin I, Searle BM, Desposito F (November 1984). "Gene dosage studies supporting localization of the structural gene for galactose-1-phosphate uridyl transferase (GALT) to band p13 of chromosome 9". American Journal of Medical Genetics. 19 (3): 539–43. doi:10.1002/ajmg.1320190316. PMID6095663.
Ashino J, Okano Y, Suyama I, Yamazaki T, Yoshino M, Furuyama J, Lin HC, Reichardt JK, Isshiki G (1995). "Molecular characterization of galactosemia (type 1) mutations in Japanese". Human Mutation. 6 (1): 36–43. doi:10.1002/humu.1380060108. PMID7550229. S2CID23500152.
Elsas LJ, Langley S, Paulk EM, Hjelm LN, Dembure PP (1995). "A molecular approach to galactosemia". European Journal of Pediatrics. 154 (7 Suppl 2): S21-7. doi:10.1007/BF02143798. PMID7671959. S2CID11937698.
Davit-Spraul A, Pourci ML, Ng KH, Soni T, Lemonnier A (November 1994). "Regulatory effects of galactose on galactose-1-phosphate uridyltransferase activity on human hepatoblastoma HepG2 cells". FEBS Letters. 354 (2): 232–6. doi:10.1016/0014-5793(94)01133-8. PMID7957929. S2CID45242645.
Lin HC, Kirby LT, Ng WG, Reichardt JK (February 1994). "On the molecular nature of the Duarte variant of galactose-1-phosphate uridyl transferase (GALT)". Human Genetics. 93 (2): 167–9. doi:10.1007/BF00210604. PMID8112740. S2CID42558872.
