The relationship between structure and function in natural surfactant proteins

Surfactant activity is a property more commonly associated with small molecules than biological macromolecules. However, the significant advantages of improved biodegradability and biocompatibility that could be presented by natural surfactant proteins has elevated interest in a group of only a few proteins where intrinsic surfactant activity appears to be the primary function. Two examples of this group, ranaspumin-2 (Rsn-2) from the foam nests of the tungara frog and latherin, the surface active protein component of horse sweat, appear to be different from other surfactant proteins in the form of their activity. However, the exact molecular basis of this activity is poorly understood. This thesis describes work to rationalise surface activity and related properties in these unusual proteins. The properties of Rsn-2 and latherin including surface activity, interaction with lipid membranes and behaviour in solution were investigated to provide further insight into the characteristics that distinguish the surfactant proteins from both conventional surfactants and other proteins. A second protein component of the foam nests, ranaspumin-1 (Rsn-1), of previously unknown function was also found to be highly surface active and is proposed to function in a similar manner to Rsn-2. A model whereby Rsn-2 functions via a clamshell-like opening was tested through a combination of specialised NMR techniques and site-directed mutagenesis. The results identified features associated with surfactant activity, all of which were consistent with the model. The potential of Rsn-2 as a recombinant fusion partner for the production of functional surfactants or foams was proven by construction of a fluorescent conjugate. Solution state NMR was used to determine the structure of latherin. Information on the dynamic processes taking place in the molecule were derived by analysis of NMR relaxation data. The structure revealed is a super roll fold, similar to a single domain of the BPI-like proteins. A model is proposed whereby latherin recognises the air-water interface via three dynamic, hydrophobic loops at one end of its long cylindrical structure and then unfolds to expose its hydrophobic core at the air-water interface

Frog foams and natural protein surfactants

Foams and surfactants are relatively rare in biology because of their potential to harm cell membranes and other delicate tissues. However, in recent work we have identified and characterized a number of natural surfactant proteins found in the foam nests of tropical frogs and other unusual sources. These proteins, and their associated foams, are relatively stable and bio-compatible, but with intriguing molecular structures that reveal a new class of surfactant activity. Here we review the structures and functional mechanisms of some of these proteins as revealed by experiments involving a range of biophysical and biochemical techniques, with additional mechanistic support coming from more recent site-directed mutagenesis studies

The structure of latherin, a surfactant allergen protein from horse sweat and saliva

Latherin is a highly surface-active allergen protein found in the sweat and saliva of horses and other equids. Its surfactant activity is intrinsic to the protein in its native form, and is manifest without associated lipids or glycosylation. Latherin probably functions as a wetting agent in evaporative cooling in horses, but it may also assist in mastication of fibrous food as well as inhibition of microbial biofilms. It is a member of the PLUNC family of proteins abundant in the oral cavity and saliva of mammals, one of which has also been shown to be a surfactant and capable of disrupting microbial biofilms. How these proteins work as surfactants while remaining soluble and cell membrane-compatible is not known. Nor have their structures previously been reported. We have used protein nuclear magnetic resonance spectroscopy to determine the conformation and dynamics of latherin in aqueous solution. The protein is a monomer in solution with a slightly curved cylindrical structure exhibiting a ‘super-roll’ motif comprising a four-stranded anti-parallel β-sheet and two opposing α-helices which twist along the long axis of the cylinder. One end of the molecule has prominent, flexible loops that contain a number of apolar amino acid side chains. This, together with previous biophysical observations, leads us to a plausible mechanism for surfactant activity in which the molecule is first localized to the non-polar interface via these loops, and then unfolds and flattens to expose its hydrophobic interior to the air or non-polar surface. Intrinsically surface-active proteins are relatively rare in nature, and this is the first structure of such a protein from mammals to be reported. Both its conformation and proposed method of action are different from other, non-mammalian surfactant proteins investigated so far

Aqueous solubilization of C60 fullerene by natural protein surfactants, latherin and ranaspumin-2

C60 fullerene is not soluble in water and dispersion usually requires organic solvents, sonication or vigorous mechanical mixing. However, we show here that mixing of pristine C60 in water with natural surfactant proteins latherin and ranaspumin-2 (Rsn-2) at low concentrations yields stable aqueous dispersions with spectroscopic properties similar to those previously obtained by more vigorous methods. Particle sizes are significantly smaller than those achieved by mechanical dispersion alone, and concentrations are compatible with clusters approximating 1:1 protein:C60 stoichiometry. These proteins can also be adsorbed onto more intractable carbon nanotubes. This promises to be a convenient way to interface a range of hydrophobic nanoparticles and related materials with biological macromolecules, with potential to exploit the versatility of recombinant protein engineering in the development of nano-bio interface devices. It also has potential consequences for toxicological aspects of these and similar nanoparticles

Resonance assignments for latherin, a natural surfactant protein from horse sweat

Latherin is an intrinsically surfactant protein of ~23 kDa found in the sweat and saliva of horses. Its function is probably to enhance the translocation of sweat water from the skin to the surface of the pelt for evaporative cooling. Its role in saliva may be to enhance the wetting, softening and maceration of the dry, fibrous food for which equines are adapted. Latherin is unusual in its relatively high content of aliphatic amino acids (~25 % leucines) that might contribute to its surfactant properties. Latherin is related to the palate, lung, and nasal epithelium carcinoma-associated proteins (PLUNCs) of mammals, at least one of which is now known to exhibit similar surfactant activity to latherin. No structures of any PLUNC protein are currently available. 15N,13C-labelled recombinant latherin was produced in Escherichia coli, and essentially all of the resonances were assigned despite the signal overlap due to the preponderance of leucines. The most notable exceptions include a number of residues located in an apparently dynamic loop region between residues 145 and 154. The assignments have been deposited with BMRB accession number 19067

Fusing Quantitative Requirements Analysis with Model-based Systems Engineering

A vision is presented for fusing quantitative requirements analysis with model-based systems engineering. This vision draws upon and combines emergent themes in the engineering milieu. “Requirements engineering” provides means to explicitly represent requirements (both functional and non-functional) as constraints and preferences on acceptable solutions, and emphasizes early-lifecycle review, analysis and verification of design and development plans. “Design by shopping” emphasizes revealing the space of options available from which to choose (without presuming that all selection criteria have previously been elicited), and provides means to make understandable the range of choices and their ramifications. “Model-based engineering” emphasizes the goal of utilizing a formal representation of all aspects of system design, from development through operations, and provides powerful tool suites that support the practical application of these principles. A first step prototype towards this vision is described, embodying the key capabilities. Illustrations, implications, further challenges and opportunities are outlined

1949 Ruby Yearbook

A digitized copy of the 1949 Ruby, the Ursinus College yearbook.https://digitalcommons.ursinus.edu/ruby/1051/thumbnail.jp

Mantle Degassing Lifetimes through Galactic Time and the Maximum Age Stagnant-lid Rocky Exoplanets can Support Temperate Climates

The ideal exoplanets to search for life are those within a star's habitable zone. However, even within the habitable zone planets can still develop uninhabitable climate states. Sustaining a temperate climate over geologic ($\sim$Gyr) timescales requires a planet contain sufficient internal energy to power a planetary-scale carbon cycle. A major component of a rocky planet's energy budget is the heat produced by the decay of radioactive elements, especially $^{40}$K, $^{232}$Th, $^{235}$U and $^{238}$U. As the planet ages and these elements decay, this radiogenic energy source dwindles. Here we estimate the probability distribution of the amount of these heat producing elements (HPEs) that enter into rocky exoplanets through Galactic history, by combining the system-to-system variation seen in stellar abundance data with the results from Galactic chemical evolution models. Using these distributions, we perform Monte-Carlo thermal evolution models that maximize the mantle cooling rate. This allows us to create a pessimistic estimate of lifetime a rocky, stagnant-lid exoplanet can support a global carbon cycle and temperate climate as a function of its mass and when it in Galactic history. We apply this framework to a sample of 17 likely rocky exoplanets with measured ages, 7 of which we predict are likely to be actively degassing today despite our pessimistic assumptions. For the remaining planets, including those orbiting TRAPPIST-1, we cannot confidently assume they currently contain sufficient internal heat to support mantle degassing at a rate sufficient to sustain a global carbon cycle or temperate climate without additional tidal heating or undergoing plate tectonics.Comment: Accepted to ApJ Letter