Polymorphism of human Alpha class glutathione transferases

Area of Science:

• Genetics
• Molecular Biology
• Biochemistry

Background:

• Single nucleotide polymorphisms (SNPs) are crucial genetic markers.
• Identifying SNPs efficiently is essential for genetic research.
• Expressed Sequence Tag (EST) databases offer a valuable resource for SNP discovery.

Purpose of the Study:

• To develop and validate a rapid method for identifying genetic polymorphisms.
• To investigate polymorphisms within the Alpha class glutathione transferase (GST) genes, specifically GSTA1 and GSTA2.
• To characterize the functional and structural impact of identified GSTA2 variants.

Main Methods:

• Screening of the EST database for polymorphisms in GSTA1 and GSTA2 genes.
• Sequence analysis for verification of putative polymorphisms.
• Polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) analysis to confirm variants in diverse populations.
• Expression, purification, and enzymatic characterization of variant GSTA2 isoforms.
• Three-dimensional structural modeling of GSTA2 polymorphisms.

Main Results:

• Ten putative polymorphisms were identified in the coding regions of GSTA1 and GSTA2 via EST database screening.
• Six polymorphisms were confirmed by sequence analysis.
• Three variants were validated using PCR/RFLP: a silent substitution in GSTA1 (K125K) and two in GSTA2 (T112S, E210A).
• Variant GSTA2 isoforms showed no significant alteration in structure or enzymatic function compared to wild-type.

Conclusions:

• EST database analysis is an efficient strategy for identifying genetic polymorphisms.
• Identified polymorphisms in GSTA1 and GSTA2 genes do not significantly affect protein function.
• This study validates a rapid approach for variant protein discovery.