Production of soluble, active acetyl serotonin methyl transferase in Leishmania tarentolae.
N-acetyl serotonin methyl transferase (ASMT) is the last enzyme in the melatonin synthesis pathway. Evidence linking autism-related disorders with disorders of melatonin metabolism, and, more specifically, with mutations of the gene encoding ASMT, prompted us to investigate the properties and localization of this enzyme. As a first step, we undertook to overproduce the protein in a recombinant host. Early attempts to produce ASMT in recombinant Escherichia coli yielded only insoluble and heavily degraded material. However, recombinant ASMT (rASMT) could be produced in soluble, active form and purified in milligram amounts when the gene was cloned and expressed in Leishmania tarentolae.
Ben-Abdallah
Mariem
M
Institut Pasteur, Platform 5 Production of Recombinant Proteins and Antibodies, 25-28, rue du Dr. Roux, 75724 Paris cedex 15, France. mariem.ben-abdallah@pasteur.fr
Bondet
Vincent
V
Fauchereau
Fabien
F
Béguin
Pierre
P
Goubran-Botros
Hany
H
Pagan
Cécile
C
Bourgeron
Thomas
T
Bellalou
Jacques
J
D000121:Acetylserotonin O-Methyltransferase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D015870:Gene Expression; D006801:Humans; D007891:Leishmania / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility
D011994:Recombinant Proteins; C523771:ASMT protein, human; D000121:Acetylserotonin O-Methyltransferase