Purification and structural characterization of a novel bacterial LMF1 homolog

Undergraduates: Lin Cao, Cassadra Hayne

Faculty Advisor: Saskia Neher
Department: Biology

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Proper functioning of many human lipases requires assistance from

the protein Lipase Maturation Factor 1 (LMF1). The exact

mechanism by which LMF1 functions is still under study, and LMF1

is also found in cells and organisms that do not secrete lipases,

suggesting it has additional, unknown functions. Because no LMF1

homologs have known structures, resolving the structure of any

protein in LMF1¿¿¿s family will offer insight into how this important

protein works, as function can be inferred from structure.

Homologs from Acidothermus cellulolyticus and Streptomyces

avermitilis bacteria, both 40% identical to human LMF1, will be

studied. Their expression was optimized by testing a variety of cell

lines and growth conditions. Protein purification strategies using a

variety of detergents and nickel-NTA columns were developed and

binding, wash, and elution buffers optimized. Subsequent work will

involve characterizing the three dimensional structures of the

homologs using macromolecular crystallography.