Identifying optimal lipid raft characteristics required to promote nanoscale protein-protein interactions on the plasma membrane

The dynamic lateral segregation of signaling proteins into microdomains is proposed to facilitate signal transduction, but the constraints on microdomain size, mobility, and diffusion that might realize this function are undefined. Here we interrogate a stochastic spatial model of the plasma membrane to determine how microdomains affect protein dynamics. Taking lipid rafts as representative microdomains, we show that reduced protein mobility in rafts segregates dynamically partitioning proteins, but the equilibrium concentration is largely independent of raft size and mobility. Rafts weakly impede small-scale protein diffusion but more strongly impede long-range protein mobility. The long-range mobility of raft-partitioning and raft-excluded proteins, however, is reduced to a similar extent. Dynamic partitioning into rafts increases specific interprotein collision rates, but to maximize this critical, biologically relevant function, rafts must be small (diameter, 6 to 14 nm) and mobile. Intermolecular collisions can also be favored by the selective capture and exclusion of proteins by rafts, although this mechanism is generally less efficient than simple dynamic partitioning. Generalizing these results, we conclude that microdomains can readily operate as protein concentrators or isolators but there appear to be significant constraints on size and mobility if microdomains are also required to function as reaction chambers that facilitate nanoscale protein-protein interactions. These results may have significant implications for the many signaling cascades that are scaffolded or assembled in plasma membrane microdomains.

CDKN2A mutation in a non-FAMMM kindred with cancers at multiple sites results in a functionally abnormal protein

The CDKN2A gene encodes p16 (CDKN2A), a cell-cycle inhibitor protein which prevents inappropriate cell cycling and, hence, proliferation. Germ-line mutations in CDKN2A predispose to the familial atypical multiple-mole melanoma (FAMMM) syndrome but also have been seen in rare families in which only 1 or 2 individuals are affected by cutaneous malignant melanoma (CMM). We therefore sequenced exons 1alpha and 2 of CDKN2A using lymphocyte DNA isolated from index cases from 67 families with cancers at multiple sites, where the patterns of cancer did not resemble those attributable to known genes such as hMLH1, hMLH2, BRCA1, BRCA2, TP53 or other cancer susceptibility genes. We found one mutation, a mis-sense mutation resulting in a methionine to isoleucine change at codon 53 (M531) of exon 2. The individual tested had developed 2 CMMs but had no dysplastic nevi and lacked a family history of dysplastic nevi or CMM. Other family members had been diagnosed with oral cancer (2 persons), bladder cancer (1 person) and possibly gall-bladder cancer. While this mutation has been reported in Australian and North American melanoma kindreds, we did not observe it in 618 chromosomes from Scottish and Canadian controls. Functional studies revealed that the CDKN2A variant carrying the M531 change was unable to bind effectively to CDK4, showing that this mutation is of pathological significance. Our results have confirmed that CDKN2A mutations are not limited to FAMMM kindreds but also demonstrate that multi-site cancer families without melanoma are very unlikely to contain CDKN2A mutations.

Estimating the loading condition of a diesel engine using instantaneous angular speed analysis

Continuing monitoring of diesel engine performance is critical for early detection of fault developments in the engine before they materialize and become a functional failure. Instantaneous crank angular speed (IAS) analysis is one of a few non intrusive condition monitoring techniques that can be utilized for such tasks. In this experimental study, IAS analysis was employed to estimate the loading condition of a 4-stroke 4-cylinder diesel engine in a laboratory condition. It was shown that IAS analysis can provide useful information about engine speed variation caused by the changing piston momentum and crankshaft acceleration during the engine combustion process. It was also found that the major order component of the IAS spectrum directly associated with the engine firing frequency (at twice the mean shaft revolution speed) can be utilized to estimate the engine loading condition regardless of whether the engine is operating at normal running conditions or in a simulated faulty injector case. The amplitude of this order component follows a clear exponential curve as the loading condition changes. A mathematical relationship was established for the estimation of the engine power output based on the amplitude of the major order component of the measured IAS spectrum.

Estimating the loading condition of a diesel engine using instantaneous angular speed analysis

Continuing monitoring of diesel engine performance is critical for early detection of fault developments in the engine before they materialize and become a functional failure. Instantaneous crank angular speed (IAS) analysis is one of a few non intrusive condition monitoring techniques that can be utilized for such tasks. In this experimental study, IAS analysis was employed to estimate the loading condition of a 4-stroke 4-cylinder diesel engine in a laboratory condition. It was shown that IAS analysis can provide useful information about engine speed variation caused by the changing piston momentum and crankshaft acceleration during the engine combustion process. It was also found that the major order component of the IAS spectrum directly associated with the engine firing frequency (at twice the mean shaft revolution speed) can be utilized to estimate the engine loading condition regardless of whether the engine is operating at normal running conditions or in a simulated faulty injector case. The amplitude of this order component follows a clear exponential curve as the loading condition changes. A mathematical relationship was established for the estimation of the engine power output based on the amplitude of the major order component of the measured IAS spectrum.