Cloning of a complete cDNA encoding human aromatase: immunochemical identification and sequence analysis

Area of Science:

• Biochemistry
• Molecular Biology
• Enzymology

Background:

• Aromatase is a key enzyme in estrogen biosynthesis.
• Understanding aromatase's molecular structure is crucial for studying its function and regulation.
• Previous characterization of aromatase at the molecular level was incomplete.

Purpose of the Study:

• To isolate and characterize the complete cDNA clone encoding human aromatase.
• To confirm the identity and function of the isolated clone.
• To analyze the molecular structure of human aromatase and its relationship to the cytochrome P-450 superfamily.

Main Methods:

• Screening a human placental cDNA library using lambda gt11.
• Antibody preparation against the cloned polypeptide.
• Western blot analysis to confirm protein identity.
• Enzyme activity inhibition assays.
• DNA sequencing and amino acid sequence analysis.

Main Results:

• A complete cDNA clone encoding human aromatase was successfully isolated.
• The clone contains a 3030 bp sequence with specific non-coding and open reading frame regions.
• The open reading frame encodes 503 amino acids.
• Antibody validation confirmed the clone's identity as aromatase cDNA.
• Sequence analysis revealed aromatase as a unique member of the cytochrome P-450 superfamily.

Conclusions:

• The isolated cDNA clone represents human aromatase.
• Human aromatase is a distinct form within the cytochrome P-450 enzyme family.
• This molecular characterization provides a foundation for further studies on aromatase function and potential therapeutic targeting.