ALDH7A1 Antibody
Aldh7a1 antibody to consanguineous parents developed seizures shortly after birth and required treatment with multiple antiepileptic drugs. Genetic evaluation revealed a homozygous likely pathogenic variant in the ALDH7A1 gene and oral administration of pyridoxine resulted in complete cessation of seizures. This case represents the first report of pyridoxine dependency in Syria, and provides a rare opportunity to explore the role of this gene in human disease.
Recombinant human ALDH7A1 was expressed in bacterial cells with an N-terminal hexahistidine tag and an His-tagged tobacco etch virus protease recognition site. The protein was purified from the soluble fraction using Ni2+-Sepharose and Superdex S200 16/60 columns. A peptide from the N-terminal region of human ALDH7A1 was identified by mass spectrometry and a synthetic peptide matching this sequence was synthesized. Antibodies against the HA epitope and the His epitope were produced with a commercial antibody production service.
High-Quality ALDH7A1 Antibody for Reliable Detection
The recombinant human ALDH7A1 had a calculated Km of 169 mm for the substrate AASA, significantly higher than that observed in human liver homogenates (181 mm) and 2.5-fold higher than that measured for recombinant seabream ALDH7A1 partially purified from cytoplasm (20 mm). AASA binds in the polar core of the active site through electrostatic interactions with Glu121 (3.04 A) and the carboxylate form forms van der Waals interactions with Tyr468, Phe476, and Ala462 (Fig. 2 C–E).
Overexpression of ALDH7A1 in CHO-Vector and CHO-hALDH7A1_v1 clone 35 cells protected the cells from hyperosmotic stress. Colony forming and DNA fragmentation assays demonstrated that the cytoprotective effects of ALDH7A1 were mediated by its activity to convert AASA to g-glutamic semialdehyde. Moreover, ALDH7A1 overexpression inhibited COPI transport by binding to the COPI protein G172E (Fig. 1j).