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Related Experiment Videos

Sequence analysis of full length cDNA for enhancing factor/phospholipase A2

S Kadam1, C Deshpande, F Coulier

  • 1Laboratory of Genetic Engineering, Tata Memorial Centre, Mumbai, India.

Indian Journal of Experimental Biology
|September 10, 1998
PubMed
Summary

Researchers identified the complete mouse Enhancing Factor (EF) gene sequence. This protein belongs to the phospholipase A2 (PLA2) family, crucial for cellular functions and signaling pathways.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Enhancing Factor (EF) protein was first identified as an epidermal growth factor modulator in mouse small intestines.
  • Initial cDNA sequencing revealed EF as a member of the non-pancreatic phospholipase A2 (PLA2) family, marking the first report of mouse PLA2.
  • The precise function and classification of EF within the PLA2 family required further investigation.

Purpose of the Study:

  • To determine the complete cDNA sequence of the mouse EF gene.
  • To analyze the predicted amino acid sequence for functional sites.
  • To establish the phylogenetic relationship of mouse EF within the PLA2 superfamily.

Main Methods:

  • Rapid Amplification of cDNA Ends (RAcE-PCR) was employed to obtain the 5' sequence of the EF gene.

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  • Computer analysis was used to predict functional sites within the amino acid sequence.
  • Phylogenetic tree inference was performed using the complete protein sequence of EF and 16 aligned sequences.

    • Main Results:

    • The complete cDNA sequence of the mouse EF gene was successfully obtained.
    • Putative sites for enzyme action, calcium binding, and heparin binding were identified in the predicted amino acid sequence.
    • Phylogenetic analysis strongly grouped mouse EF with other membrane-associated PLA2 enzymes (98% bootstrap value).

    Conclusions:

    • The Enhancing Factor (EF) protein is confirmed as a mouse phospholipase A2 (PLA2) enzyme.
    • The identified functional sites provide insights into EF's catalytic and binding activities.
    • Mouse EF is evolutionarily classified as a membrane-associated PLA2, contributing to our understanding of this enzyme family.