Our main research interest is defining the reaction and regulatory mechanisms of the first and terminal heme biosynthetic pathway enzymes, 5-aminolevulinate synthase (ALAS) and ferrochelatase (FC). Iron overload is a clinically important feature of sideroblastic anemia, X-linked SA, and myelodysplastic syndrome, which often results from either ineffective hematopoiesis or the repeated transfusions undergone by the patients to manage their erythropoietic defects. The pathological consequences of mitochondrial mishandling of iron and heme synthesis are also evident in erythropoietic porphyrias. There is no cure for the above disorders, and thus understanding the mechanisms of the terminal stages of erythropoiesis becomes necessary for discovering novel therapeutic targets. Towards this goal, our on-going research focuses on establishing 1) whether succinyl-CoA synthetase b-subunit allosterically fine-tunes the activity of erythroid ALAS and 2) the mechanism of Fe2+ delivery to FC.
Awards/Honors
Elected Member (Nominating Committee of the Division of Biological Chemistry, American Chemical Society 2002 - 2004)
Elected Vice-Chair (Year 2000) and Chair (Year 2002) (Gordon Research Conference, The Chemistry and Biology of Tetrapyrroles - 2000)
National Science Foundation Young Investigator Award (1992 - 1997)
Presidential Young Faculty Award (University of South Florida 1995 - 1996)
Memberships
Member (American Society for Biochemistry and Molecular Biology, 2007 - Present)
Member (Biophysical Society, 2007 - Present)
Member (Protein Society, 2007 - Present)
Member (American Association for Advancement of Science, 2007 - Present)
Member (Sociedade Portuguesa de Bioquimica, 2007 - Present)
Member (American Society for Hematology, 2007 - Present)
Member (American Porphyria Foundation, 2007 - Present)
Member (Society of Porphyrins & Phatlocyanines, 2007 - Present)
Member (American Chemical Society, 2007 - Present)
Recent Publications
Tan D, Harrison T, Hunter GA, Ferreira GC. Role of arginine 439 in substrate binding of 5-aminolevulinate synthase.Biochemistry. 37(6) : 1478-84, 1998.
Al-Karadaghi, Salam|Franco, Ricardo|Hansson, Mats|Shelnutt, John A.|Isaya, Grazia|Ferreira, Gloria C. Chelatases: distort to select?Trends in Biochemical Sciences. 31(3) : 135-42, 2006.
Fratz-Berilla EJ, Breydo L, Gouya L, Puy H, Uversky VN, Ferreira GC. Isoniazid inhibits human erythroid 5-aminolevulinate synthase: Molecular mechanism and tolerance study with four X-linked protoporphyria patients.Biochimica et biophysica acta. 1863(2) : 428-439, 2017.
Na I, DeForte S, Stojanovski BM, Ferreira GC, Uversky VN. Molecular dynamics analysis of the structural and dynamic properties of the functionally enhanced hepta-variant of mouse 5-aminolevulinate synthase.Journal of biomolecular structure & dynamics. : 1-14, 2017.
Stojanovski BM Breydo L Uversky VN Ferreira GC. Macromolecular Crowders and Osmolytes Modulate the Structural and Catalytic Properties of Alkaline Molten Globular 5-Aminolevulinate Synthase.RSC Advances,. 6: 114541-114552; doi: 10.1039/C6RA22533, 2016.
Stojanovski BM, Breydo L, Uversky VN, Ferreira GC. The unfolding pathways of the native and molten globule states of 5-aminolevulinate synthase.Biochemical and biophysical research communications. 480(3) : 321-327, 2016.
Hunter GA Vankayala SL Gillam ME Kearns F Woodcock HL and Ferreira
GC. The conserved active site histidine-glutamate pair of ferrochelatase
coordinately catalyzes porphyrin metalation.J. Porphyrins & Phthalocyanines. 20(20) : 556-569, 2016.
Tan D, Barber MJ, Ferreira GC. The role of tyrosine 121 in cofactor binding of 5-aminolevulinate synthase.Protein science : a publication of the Protein Society. 7(5) : 1208-13, 1998.
Stojanovski BM, Breydo L, Uversky VN, Ferreira GC. Murine erythroid 5-aminolevulinate synthase: Truncation of a disordered N-terminal extension is not detrimental for catalysis.Biochimica et Biophysica Acta. 1864(5) : 441-52, 2016.
Tan D, Ferreira GC. Active site of 5-aminolevulinate synthase resides at the subunit interface. Evidence from in vivo heterodimer formation.Biochemistry. 35(27) : 8934-41, 1996.
Gong J, Kay CJ, Barber MJ, Ferreira GC. Mutations at a glycine loop in aminolevulinate synthase affect pyridoxal phosphate cofactor binding and catalysis.Biochemistry. 35(45) : 14109-17, 1996.
Söderberg C, Gillam ME, Ahlgren EC, Hunter GA, Gakh O, Isaya G, Ferreira GC, Al-Karadaghi S. The Structure of the Complex between Yeast Frataxin and Ferrochelatase: CHARACTERIZATION AND PRE-STEADY STATE REACTION OF FERROUS IRON DELIVERY AND HEME SYNTHESIS.The Journal of Biological Chemistry. 291(22) : 11887-98, 2016.
Stojanovski BM, Ferreira GC. Asn-150 of Murine Erythroid 5-Aminolevulinate Synthase Modulates the Catalytic Balance between the Rates of the Reversible Reaction.The Journal of Biological Chemistry. 290(52) : 30750-61, 2015.
Ferreira GC. Ferrochelatase binds the iron-responsive element present in the erythroid 5-aminolevulinate synthase mRNA.Biochemical and biophysical research communications. 214(3) : 875-8, 1995.
Franco R, Moura JJ, Moura I, Lloyd SG, Huynh BH, Forbes WS, Ferreira GC. Characterization of the iron-binding site in mammalian ferrochelatase by kinetic and Mössbauer methods.The Journal of biological chemistry. 270(44) : 26352-7, 1995.
Stojanovski BM, Ferreira GC. Murine erythroid 5-aminolevulinate synthase: Adenosyl-binding site Lys221 modulates substrate binding and catalysis.FEBS Open Bio. 5: 824-31, 2015.
Gong J, Ferreira GC. Aminolevulinate synthase: functionally important residues at a glycine loop, a putative pyridoxal phosphate cofactor-binding site.Biochemistry. 34(5) : 1678-85, 1995.
Fratz EJ, Hunter GA, Ferreira GC. Expression of murine 5-aminolevulinate synthase variants causes protoporphyrin IX accumulation and light-induced mammalian cell death.PloS One. 9(4) : e93078, 2014.
Ferreira GC. Heme biosynthesis: biochemistry, molecular biology, and relationship to disease.Journal of bioenergetics and biomembranes. 27(2) : 147-50, 1995.
Stojanovski BM, Hunter GA, Jahn M, Jahn D, Ferreira GC. Unstable reaction intermediates and hysteresis during the catalytic cycle of 5-aminolevulinate synthase: implications from using pseudo and alternate substrates and a promiscuous enzyme variant.The Journal of Biological Chemistry. 289(33) : 22915-25, 2014.
Ferreira GC, Gong J. 5-Aminolevulinate synthase and the first step of heme biosynthesis.Journal of bioenergetics and biomembranes. 27(2) : 151-9, 1995.
Stojanovski BM, Breydo L, Hunter GA, Uversky VN, Ferreira GC. Catalytically active alkaline molten globular enzyme: Effect of pH and temperature on the structural integrity of 5-aminolevulinate synthase.Biochimica et Biophysica Acta. 1844(12) : 2145-54, 2014.
Hunter GA, Ferreira GC. A continuous spectrophotometric assay for 5-aminolevulinate synthase that utilizes substrate cycling.Analytical biochemistry. 226(2) : 221-4, 1995.
Hunter GA, Ferreira GC. Molecular enzymology of 5-Aminolevulinate synthase, the gatekeeper of heme biosynthesis.Biochimica Biophysica Acta. 1814(11) : 1467-73, 2011.
Ferreira GC, Franco R, Lloyd SG, Moura I, Moura JJ, Huynh BH. Structure and function of ferrochelatase.Journal of bioenergetics and biomembranes. 27(2) : 221-9, 1995.
Ferreira GC, Vajapey U, Hafez O, Hunter GA, Barber MJ. Aminolevulinate synthase: lysine 313 is not essential for binding the pyridoxal phosphate cofactor but is essential for catalysis.Protein science : a publication of the Protein Society. 4(5) : 1001-6, 1995.
Ferreira GC. Mammalian ferrochelatase. Overexpression in Escherichia coli as a soluble protein, purification and characterization.The Journal of biological chemistry. 269(6) : 4396-400, 1994.
Hunter GA, Al-Karadaghi S, Ferreira GC. FERROCHELATASE: THE CONVERGENCE OF THE PORPHYRIN BIOSYNTHESIS AND IRON TRANSPORT PATHWAYS.Journal of porphyrins and phthalocyanines. 15(5-6) : 350-356, 2011.
Ferreira GC, Franco R, Lloyd SG, Pereira AS, Moura I, Moura JJ, Huynh BH. Mammalian ferrochelatase, a new addition to the metalloenzyme family.The Journal of biological chemistry. 269(10) : 7062-5, 1994.
Franco R, Al-Karadaghi S, Ferreira GC. Resonance Raman Spectroscopic Examination of Ferrochelatase-induced Porphyrin Distortion.Journal of Porphyrins and Phthalocyanines. 15(5) : 357-363, 2011.
Turbeville TD, Zhang J, Adams WC, Hunter GA, Ferreira GC. Functional asymmetry for the active sites of linked 5-aminolevulinate synthase and 8-amino-7-oxononanoate synthase.Archives of biochemistry and biophysics. 511(1-2) : 107-17, 2011.
Jabs EW, Thomas PJ, Bernstein M, Coss C, Ferreira GC, Pedersen PL. Chromosomal localization of genes required for the terminal steps of oxidative metabolism: alpha and gamma subunits of ATP synthase and the phosphate carrier.Human genetics. 93(5) : 600-2, 1994.
McIntyre NR Franco R Shelnutt JA and Ferreira GC. Nickel(II) Chelatase
Variants Directly Evolved from Murine Ferrochelatase: Porphyrin Distortion and Kinetic
Mechanism.Biochemistry. 50: 1535-1544, 2011.
Ferreira GC, Neame PJ, Dailey HA. Heme biosynthesis in mammalian systems: evidence of a Schiff base linkage between the pyridoxal 5''-phosphate cofactor and a lysine residue in 5-aminolevulinate synthase.Protein science : a publication of the Protein Society. 2(11) : 1959-65, 1993.
To-Figueras J, Ducamp S, Clayton J, Badenas C, Delaby C, Ged C, Lyoumi S, Gouya L, de Verneuil H, Beaumont C, Ferreira GC, Deybach JC, Herrero C, Puy H. ALAS2 acts as a modifier gene in patients with congenital erythropoietic porphyria.Blood. 118(6) : 1443-51, 2011.
Ferreira GC, Pedersen PL. Phosphate transport in mitochondria: past accomplishments, present problems, and future challenges.Journal of bioenergetics and biomembranes. 25(5) : 483-92, 1993.
Ferreira GC, Dailey HA. Expression of mammalian 5-aminolevulinate synthase in Escherichia coli. Overproduction, purification, and characterization.The Journal of biological chemistry. 268(1) : 584-90, 1993.
Ferreira GC. Erythroid 5-aminolevulinate synthase and X-linked sideroblastic anemia.The Journal of the Florida Medical Association. 80(7) : 481-3, 1993.
Lendrihas T Hunter GA and Ferreira GC. Role of Serine-254 in
Conformational Switching of Murine 5-Aminolevulinate Synthase.J. Biol. Chem.. 285: 3351-3359, 2010.
Ferreira GC, Pedersen PL. Overexpression of higher eukaryotic membrane proteins in bacteria. Novel insights obtained with the liver mitochondrial proton/phosphate symporter.The Journal of biological chemistry. 267(8) : 5460-6, 1992.
Lendrihas T, Hunter GA, Ferreira GC. Targeting the active site gate to yield hyperactive variants of 5-aminolevulinate synthase.The Journal of Biological Chemistry. 285(18) : 13704-11, 2010.
Hunter GA, Ferreira GC. Identification and characterization of an inhibitory metal ion-binding site in ferrochelatase.The Journal of Biological Chemistry. 285(53) : 41836-42, 2010.
Pratt RD, Ferreira GC, Pedersen PL. Mitochondrial phosphate transport. Import of the H+/Pi symporter and role of the presequence.The Journal of biological chemistry. 266(2) : 1276-80, 1991.
Lendrihas T, Zhang J, Hunter GA, Ferreira GC. Arg-85 and Thr-430 in murine 5-aminolevulinate synthase coordinate acyl-CoA-binding and contribute to substrate specificity.Protein science : a publication of the Protein Society. 18(9) : 1847-59, 2009.
Ferreira GC, Pratt RD, Pedersen PL. Mitochondrial proton/phosphate transporter. An antibody directed against the COOH terminus and proteolytic cleavage experiments provides new insights about its membrane topology.The Journal of biological chemistry. 265(34) : 21202-6, 1990.
Hunter GA, Ferreira GC. 5-aminolevulinate synthase: catalysis of the first step of heme biosynthesis.Cellular and Molecular Biology (Noisy-le-Grand, France). 55(1) : 102-10, 2009.
Ferreira GC, Pratt RD, Pedersen PL. Energy-linked anion transport. Cloning, sequencing, and characterization of a full length cDNA encoding the rat liver mitochondrial proton/phosphate symporter.The Journal of biological chemistry. 264(26) : 15628-33, 1989.
Karlberg T, Hansson MD, Yengo RK, Johansson R, Thorvaldsen HO, Ferreira GC, Hansson M, Al-Karadaghi S. Porphyrin binding and distortion and substrate specificity in the ferrochelatase reaction: the role of active site residues.Journal of Molecular Biology. 378(5) : 1074-83, 2008.
Ferreira GC, Andrew TL, Karr SW, Dailey HA. Organization of the terminal two enzymes of the heme biosynthetic pathway. Orientation of protoporphyrinogen oxidase and evidence for a membrane complex.The Journal of biological chemistry. 263(8) : 3835-9, 1988.
Hunter GA, Sampson MP, Ferreira GC. Metal ion substrate inhibition of ferrochelatase.The Journal of Biological Chemistry. 283(35) : 23685-91, 2008.
Ferreira GC, Dailey HA. Mouse protoporphyrinogen oxidase. Kinetic parameters and demonstration of inhibition by bilirubin.The Biochemical journal. 250(2) : 597-603, 1988.
Hunter GA, Zhang J, Ferreira GC. Transient kinetic studies support refinements to the chemical and kinetic mechanisms of aminolevulinate synthase.The Journal of Biological Chemistry. 282(32) : 23025-35, 2007.
Ferreira GC, Dailey HA. Reconstitution of the two terminal enzymes of the heme biosynthetic pathway into phospholipid vesicles.The Journal of biological chemistry. 262(9) : 4407-12, 1987.
Rigler MW, Ferreira GC, Patton JS. Intramembranous particles are clustered on microvillus membrane vesicles.Biochimica et biophysica acta. 816(1) : 131-41, 1985.
Gakh O, Park S, Liu G, Macomber L, Imlay JA, Ferreira GC, Isaya G. Mitochondrial iron detoxification is a primary function of frataxin that limits oxidative damage and preserves cell longevity.Human Molecular Genetics. 15(3) : 467-79, 2006.
Shi Z, Franco R, Haddad R, Shelnutt JA, Ferreira GC. The conserved active-site loop residues of ferrochelatase induce porphyrin conformational changes necessary for catalysis.Biochemistry. 45(9) : 2904-12, 2006.
Dias JS, Macedo AL, Ferreira GC, Peterson FC, Volkman BF, Goodfellow BJ. The first structure from the SOUL/HBP family of heme-binding proteins, murine P22HBP.The Journal of biological chemistry. 281(42) : 31553-61, 2006.
Shi Z, Ferreira GC. Modulation of inhibition of ferrochelatase by N-methylprotoporphyrin.The Biochemical Journal. 399(1) : 21-8, 2006.
Franco R, Bai G, Prosinecki V, Abrunhosa F, Ferreira GC, Bastos M. Porphyrin-substrate binding to murine ferrochelatase: effect on the thermal stability of the enzyme.The Biochemical journal. 386(Pt 3) : 599-605, 2005.
Zhang J, Cheltsov AV, Ferreira GC. Conversion of 5-aminolevulinate synthase into a more active enzyme by linking the two subunits: spectroscopic and kinetic properties.Protein science : a publication of the Protein Society. 14(5) : 1190-200, 2005.
Hunter GA, Rivera E, Ferreira GC. Supraphysiological concentrations of 5-aminolevulinic acid dimerize in solution to produce superoxide radical anions via a protonated dihydropyrazine intermediate.Archives of biochemistry and biophysics. 437(2) : 128-37, 2005.
Dias JS, Macedo AL, Ferreira GC, Jeanty N, Taketani S, Goodfellow BJ, Peterson FC, Volkman BF. 1H, 15N and 13C resonance assignments of the heme-binding protein murine p22HBP.Journal of biomolecular NMR. 32(4) : 338, 2005.
O''Neill HA, Gakh O, Park S, Cui J, Mooney SM, Sampson M, Ferreira GC, Isaya G. Assembly of human frataxin is a mechanism for detoxifying redox-active iron.Biochemistry. 44(2) : 537-45, 2005.
Shipovskov, S., Karlberg, T., Fodje, M., Hansson, M. D., Ferreira, G. C, Hansson, M., Reimann, C. T. and Al-Karadaghi, S. Ferrochelatase-catalyzed Metallation of the Transition-state Inhibitor N-methyl Mesoporphyrin: A Time-resolved Crystallographic and Mass Spectrometric Study.Journal of Molecular Biololgy. 352: 1081-1090, 2005.
Shi Z, Ferreira GC. Probing the active site loop motif of murine ferrochelatase by random mutagenesis.The Journal of biological chemistry. 279(19) : 19977-86, 2004.
Ferreira, G. C. Heme SynthesisEncyclopedia of Biological Chemistry. ((Lennarz, W. and Lane, M. D., Eds.), Academic Press, San Diego, CA, USA.) : 2:362-2:366, 2004.
Park S, Gakh O, O''Neill HA, Mangravita A, Nichol H, Ferreira GC, Isaya G. Yeast frataxin sequentially chaperones and stores iron by coupling protein assembly with iron oxidation.The Journal of biological chemistry. 278(33) : 31340-51, 2003.
Krishnamachary, B., Berg-Dixon, S., Kelly, B., Agani, F., Feldser, D., Ferreira, G., Iyer, N., LaRusch, J., Pak, B., Taghavi, P., and Semenza, G. L. Regulation of Colon Carcinoma Cell Invasion by Hypoxia-inducible Factor 1.Cancer Res.. 63: 1138-1143, 2003.
Cheltsov AV, Guida WC, Ferreira GC. Circular permutation of 5-aminolevulinate synthase: effect on folding, conformational stability, and structure.The Journal of biological chemistry. 278(30) : 27945-55, 2003.
Hofer T, Wenger RH, Kramer MF, Ferreira GC, Gassmann M. Hypoxic up-regulation of erythroid 5-aminolevulinate synthase.Blood. 101(1) : 348-50, 2003.
Shi Z, Ferreira GC. A continuous anaerobic fluorimetric assay for ferrochelatase by monitoring porphyrin disappearance.Analytical biochemistry. 318(1) : 18-24, 2003.
Ferreira GC, Cheltsov AV. Circular permutation of 5-aminolevulinate synthase as a tool to evaluate folding, structure and function.Cellular and molecular biology (Noisy-le-Grand, France). 48(1) : 11-6, 2002.
Zhang J, Ferreira GC. Transient state kinetic investigation of 5-aminolevulinate synthase reaction mechanism.The Journal of biological chemistry. 277(47) : 44660-9, 2002.
Ferreira GC, Zhang JS. Mechanism of 5-aminolevulinate synthase and the role of the protein environment in controlling the cofactor chemistry.Cellular and molecular biology (Noisy-le-Grand, France). 48(8) : 827-33, 2002.
Ferreira GC, Franco R, Mangravita A, George GN. Unraveling the substrate-metal binding site of ferrochelatase: an X-ray absorption spectroscopic study.Biochemistry. 41(15) : 4809-18, 2002.
Lu Y, Sousa A, Franco R, Mangravita A, Ferreira GC, Moura I, Shelnutt JA. Binding of protoporphyrin IX and metal derivatives to the active site of wild-type mouse ferrochelatase at low porphyrin-to-protein ratios.Biochemistry. 41(26) : 8253-62, 2002.
Franco R, Pereira AS, Tavares P, Mangravita A, Barber MJ, Moura I, Ferreira GC. Substitution of murine ferrochelatase glutamate-287 with glutamine or alanine leads to porphyrin substrate-bound variants.The Biochemical journal. 356(Pt 1) : 217-22, 2001.
Goodfellow BJ, Dias JS, Ferreira GC, Henklein P, Wray V, Macedo AL. The solution structure and heme binding of the presequence of murine 5-aminolevulinate synthase.FEBS letters. 505(2) : 325-31, 2001.
Cheltsov AV, Barber MJ, Ferreira GC. Circular permutation of 5-aminolevulinate synthase. Mapping the polypeptide chain to its function.The Journal of biological chemistry. 276(22) : 19141-9, 2001.
Kramer MF, Gunaratne P, Ferreira GC. Transcriptional regulation of the murine erythroid-specific 5-aminolevulinate synthase gene.Gene. 247(1-2) : 153-66, 2000.
Franco R, Ma JG, Lu Y, Ferreira GC, Shelnutt JA. Porphyrin interactions with wild-type and mutant mouse ferrochelatase.Biochemistry. 39(10) : 2517-29, 2000.
Schneider-Yin X, Gouya L, Dorsey M, Rüfenacht U, Deybach JC, Ferreira GC. Mutations in the iron-sulfur cluster ligands of the human ferrochelatase lead to erythropoietic protoporphyria.Blood. 96(4) : 1545-9, 2000.
Feldser, D., Agani, F., Iyer, N. V., Pak, B., Ferreira, G., and Semenza, G. L. Reciprocal Positive Regulation of Hypoxia-inducible Factor 1 and Insulin-like Growth Factor 2.Cancer Res.. 59: 3915-3918, 1999.
Ferreira GC. Ferrochelatase.The international journal of biochemistry & cell biology. 31(10) : 995-1000, 1999.
Hunter GA, Ferreira GC. Lysine-313 of 5-Aminolevulinate Synthase Acts as a General Base during Formation of the Quinonoid Reaction Intermediates.Biochemistry. 38(38) : 12526, 1999.
Hunter GA, Ferreira GC. Pre-steady-state reaction of 5-aminolevulinate synthase. Evidence for a rate-determining product release.The Journal of biological chemistry. 274(18) : 12222-8, 1999.
Hunter GA, Ferreira GC. Lysine-313 of 5-aminolevulinate synthase acts as a general base during formation of the quinonoid reaction intermediates.Biochemistry. 38(12) : 3711-8, 1999.
Gong J, Hunter GA, Ferreira GC. Aspartate-279 in aminolevulinate synthase affects enzyme catalysis through enhancing the function of the pyridoxal 5''-phosphate cofactor.Biochemistry. 37(10) : 3509-17, 1998.
