Expression and functional studies of a recombinant surfactant protein B

Moulton, Jonathan (2013) Expression and functional studies of a recombinant surfactant protein B. Masters thesis, Memorial University of Newfoundland.

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Abstract

Of the components of the respiratory system, one of the most crucial is the pulmonary surfactant. This mixture of phospholipids, cholesterol, and proteins act to reduce surface tension inside the lungs as well as fight infection. The surfactant associated proteins vary in their structure and function but the most important is surfactant protein B (SP-B) which is essential to life. Despite its importance, it has not yet been structurally characterized and as a result, its function still remains a mystery. Part of the problem is the difficulty in acquiring enough of the protein to work with. To attempt to solve this problem, a method has been developed to recombinantly express a variant of human SP-B in Escherichia coli in relative abundance. Also, a series of functional tests on the recombinant SP-B were completed to determine if it is a viable alternative to native SP-B for use in the laboratory and possible clinical applications. The functional tests included membrane leakage and lipid mixing assays, as well as captive bubble surfactometer studies. These experiments revealed that the recombinant SP-B displayed normal SP-B activity albeit not as effective as native SP-B.

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