Calculating Single-Channel Permeability and Conductance from Transition Paths

Permeability and conductance are the major transport properties of membrane channels, quantifying the rate of channel crossing by the solute. It is highly desirable to calculate these quantities in all-atom molecular dynamics simulations. When the solute crossing rate is low, however, direct methods would require prohibitively long simulations, and one thus typically adopts alternative strategies based on the free energy of single solute along the channel. Here we present a new method to calculate the crossing rate by initiating unbiased trajectories in which the solute is released at the free energy barrier. In this method, the total time the solute spends in the barrier region during a channel crossing (transition path) is used to determine the kinetic rate. Our method achieves a significantly higher statistical accuracy than the classical reactive flux method, especially for diffusive barrier crossing. Our test on ion permeation through a carbon nanotube verifies that the method correctly predicts the crossing rate and reproduces the spontaneous crossing events as in long equilibrium simulations. The rigorous and efficient method here will be valuable for quantitatively connecting simulations to experimental measurement of membrane channels

Radial Spokes-A Snapshot of the Motility Regulation, Assembly, and Evolution of Cilia and Flagella

Propulsive forces generated by cilia and flagella are used in events that are critical for the thriving of diverse eukaryotic organisms in their environments. Despite distinctive strokes and regulations, the majority of them adopt the 9+2 axoneme that is believed to exist in the last eukaryotic common ancestor. Only a few outliers have opted for a simpler format that forsakes the signature radial spokes and the central pair apparatus, although both are unnecessary for force generation or rhythmicity. Extensive evidence has shown that they operate as an integral system for motility control. Recent studies have made remarkable progress on the radial spoke. This review will trace how the new structural, compositional, and evolutional insights pose significant implications on flagella biology and, conversely, ciliopathy

The Noncanonical Roles of Two Primordial Molecules in Flagella

Motile cilia and flagella are ancient organelles that eukaryotic organisms today still rely on to thrive in their natural environment. Not surprisingly, accumulated evidence has shown that the intricate motility machinery, the microtubule-based axoneme, is evolutionarily conserved down to the molecular level. This notion is epitomized by the signature axonemal complex, the radial spoke (RS). The RS is part of a control center conferring the high frequency and tightly regulated movement. Key RS proteins discovered in biflagellate green alga, Chlamydomonas reinhardtii, are also generated by nearly all ciliated organisms, including Homo sapiens. Among them are two subunits from primordial protein families, nucleoside diphosphate kinase (NDK) and heat shock protein (HSP) 40, that are positioned at a critical juncture where two arms merge into a singular stalk in the Y-shaped complex. While it is well accepted that NDKs and HSP40s maintain nucleotide homeostasis and assist HSP70 chaperone in protein folding respectively, these actions fail to explain observations in myriads of vital cellular processes. Using the experimental approaches possible in the biflagellate green algae, genetics in particular, this dissertation discovers non-canonical applications of these two spoke proteins in the assembly and motility of the RS and entire flagellum. The versatility sheds light on the canonical mechanisms, the diverse processes adopting the non-canonical mechanisms, and their preservation since their conscription perhaps by the cell ancestral to all eukaryotes