Time to Face the Fats: What Can Mass Spectrometry Reveal about the Structure of Lipids and Their Interactions with Proteins?

Since the 1950s, X-ray crystallography has been the mainstay of structural biology, providing detailed atomic-level structures that continue to revolutionize our understanding of protein function. From recent advances in this discipline, a picture has emerged of intimate and specific interactions between lipids and proteins that has driven renewed interest in the structure of lipids themselves and raised intriguing questions as to the specificity and stoichiometry in lipid-protein complexes. Herein we demonstrate some of the limitations of crystallography in resolving critical structural features of ligated lipids and thus determining how these motifs impact protein binding. As a consequence, mass spectrometry must play an important and complementary role in unraveling the complexities of lipid-protein interactions. We evaluate recent advances and highlight ongoing challenges towards the twin goals of (1) complete structure elucidation of low, abundant, and structurally diverse lipids by mass spectrometry alone, and (2) assignment of stoichiometry and specificity of lipid interactions within protein complexes.     

A oxygen-dependent reductive dechlorination of chloroform by (Bisdiphenylphosphinomethane)cobalthexacarbonyl Co2(CO)6(dppm)

The reaction of Co₂(μ-dppm)(CO)₆ with aerated chloroform affords [Co{Ph₂P(O)CH₂P(O)Ph₂}₃][CoCl₄] in low yield, and this reaction is demonstrated to be prevented under anaerobic conditions representing an unusual example of a reductive dechlorination which only takes place in the presence of oxygen.     

Engineering of an "unnatural" natural product by swapping polyketide synthase domains in Aspergillus nidulans

An StcA-AfoE hybrid polyketide synthase (PKS), generated by swapping the AfoE (asperfuranone biosynthesis) SAT domain with the StcA (sterigmatocystin biosynthesis) SAT domian, produced a major new metabolite with the same chain length as the native AfoE product. Structure elucidation allowed us to propose a likely pathway, and feeding studies supported the hypothesis that the chain length of PKS metabolites may be under precise control of KS and PT domains.     

Redox-active bridging ligands based on indigo diimine ("Nindigo") derivatives

Reactions of indigo with a variety of substituted anilines produce the corresponding indigo diimines ("Nindigos") in good yields. Nindigo coordination complexes are subsequently prepared by reactions of the Nindigo ligands with Pd(hfac)(2). In most cases, binuclear complexes are obtained in which the deprotonated Nindigo bridges two Pd(hfac) moieties in the expected bis-bidentate binding mode. When the Nindigo possesses bulky substituents on the imine (mesityl, 2,6-dimethylphenyl, 2,6-diisopropylphenyl, etc.), mononuclear Pf(hfac) complexes are obtained in which the Nindigo core has isomerized from a trans- to a cis-alkene; in these structures, the palladium is bound to the cis-Nindigo ligand at the two indole nitrogen atoms; the remaining proton is bound between the imine nitrogen atoms. The palladium complexes possess intense electronic absorption bands [near 920 nm for the binuclear complexes and 820 nm for the mononuclear cis-Nindigo complexes; extinction coefficients are (1.0-2.0) × 10(4) M(-1) cm(-1)] that are ligand-centered (π-π*) transitions. Cyclic voltammetry investigations reveal multiple redox events that are also ligand-centered in origin. All of the palladium complexes can be reversibly oxidized in two sequential one-electron steps; the binuclear complexes are reduced in a two-electron process whose reversibility depends on the Nindigo ligand substituent; the mononuclear palladium species show two one-electron reductions, only the first of which is quasi-reversible.     

Simultaneous concentration and velocity field measurements in a shock-accelerated mixing layer

A novel technique to obtain simultaneous velocity and concentration measurements is applied to the Richtmyer–Meshkov instability. After acceleration by a Mach 2.2 shock wave, the interface between the two gases develops into a turbulent mixing layer. A time-separated pair of acetone planar laser-induced fluorescence images are processed to yield concentration and, through application of the Advection-Corrected Correlation Image Velocimetry technique, velocity fields. This is the first application of this technique to shock-accelerated flows. We show that when applied to numerical simulations, this technique reproduces the velocity field to a similar quality as particle image velocimetry. When applied to the turbulent mixing layer of the experiments, information about the Reynolds number and anisotropy of the flow is obtained.     

Capacitive micromachined ultrasonic transducers using commercial multi-user MUMPs process: Capability and limitations

The objective of this work is to construct capacitive micromachined ultrasound transducers (cMUTs) using multi-user microelectromechanical systems (MEMS) processess (MUMPs) and to analyze the capability of this process relative to the customized processes commonly in use. The MUMPs process has the advantages of low cost and accessibility to general users since it is not necessary to have access to customized fabrication capability such as wafer-bonding and sacrificial release processes. While other researchers have reported fabricating cMUTs using the MUMPs process none has reported the limitations in the process that arise due to the use of standard design rules that place limitations on the material thicknesses, gap thicknesses, and materials that may be used. In this paper we explain these limitations, and analyze the capabilities using 1D modeling, Finite Element Analysis, and experimental devices. We show that one of the limitations is that collapse voltage and center frequency can not be controlled independently. However, center frequencies up to 9 MHz can be achieved with collapse voltages of less than 200 V making such devices suitable for medical and non-destructive evaluation imaging applications. Since the membrane and base electrodes are made of polysilicon, there is a larger series resistance than that resulting from processes that use metal electrodes. We show that the series resistance is not a significant problem. The conductive polysilicon can also destroy the cMUT if the top membrane is pulled in the bottom. As a solution we propose the application of an additional dielectric layer. Finally we demonstrate a device built with a novel beam construction that produces transmitted pressure pulse into air with 6% bandwidth and agrees reasonably well with the 1D model. We conclude that cMUTs made with MUMPs process have some limitations that are not present in customized processes. However, these limitations may be overcome with the proper design considerations that we have presented putting a low cost, highly accessible means of making cMUT devices into the hands of academic and industrial researchers.