N-Acylphosphatidylethanolamines (NAPEs) are precursors of bioactive N-acylethanolamines, including the endocannabinoid anandamide. In animal tissues, NAPE is formed by transfer of a fatty acyl chain at the sn-1 position of glycerophospholipids to the amino group of phosphatidylethanolamine (PE), and this reaction is believed to be the principal rate-limiting step in N-acylethanolamine synthesis. However, the Ca 2؉ -dependent, membrane-associated N-acyltransferase (NAT) responsible for this reaction has not yet been cloned. In this study, on the basis of the functional similarity of NAT to lecithin-retinol acyltransferase (LRAT), we examined a possible PE N-acylation activity in two rat LRAT homologous proteins. Upon overexpression in COS-7 cells, one protein, named rat LRAT-like protein (RLP)-1, catalyzed transfer of a radioactive acyl group from phosphatidylcholine (PC) to PE, resulting in the formation of radioactive NAPE. However, the RLP-1 activity was detected mainly in the cytosolic rather than membrane fraction and was little stimulated by Ca 2؉ . Moreover, RLP-1 did not show selectivity with respect to the sn-1 and sn-2 positions of PC as an acyl donor and therefore could generate N-arachidonoyl-PE (anandamide precursor) from 2-arachidonoyl-PC and PE. In contrast, under the same assay conditions, partially purified NAT from rat brain was highly Ca 2؉ -dependent, membrane-associated, and specific for the sn-1-acyl group of PC. RLP-1 mRNA was expressed predominantly in testis among various rat tissues, and the testis cytosol exhibited an RLP-1-like activity. These results reveal that RLP-1 can function as a PE N-acyltransferase, catalytically distinguishable from the known Ca 2؉ -dependent NAT.
H-Rev107 is a protein that was previously cloned as a negative regulator of proto-oncogene Ras and classified as a class II tumor suppressor. Its structural similarity to lecithin retinol acyltransferase and Ca 21 -independent phosphatidylethanolamine (PE) N-acyltransferase led us to analyze H-Rev107 as an enzyme involved in phospholipid metabolism. Here, we show that recombinant H-Rev107s from rat, human, and mouse possess phospholipase (PL) A 1 or A 2 activity toward phosphatidylcholine (PC). Further examination with purified recombinant protein revealed that H-Rev107 functions as a cytosolic Ca 21 -independent PLA 1/2 for PC and PE with higher PLA 1 activity than PLA 2 activity. Dithiothreitol and iodoacetic acid exhibited stimulatory and inhibitory effects, respectively. Histidine-21 and cysteine-111 of rat H-Rev107 were presumed to form a catalytic dyad based on database analysis, and their single mutants were totally inactive. These results suggested that H-Rev107 is a hydrolase of the thiol type. The N-terminal proline-rich and C-terminal hydrophobic domains of H-Rev107 were earlier reported to be responsible for the regulation of cell proliferation. Analysis of deletion mutants indicated that these domains are also catalytically essential, suggesting relevance of the catalytic activity to the anti-proliferative activity.-Uyama, T., J.
Abstract. This
study investigates aerosol hygroscopicity, mixing state, and cloud
condensation nucleation as part of the Atmosphere–Aerosol–Boundary
Layer–Cloud Interaction Joint Experiment performed in the summer of 2016 at
Xingtai (XT), a suburban site located in the center of the North China Plain
(NCP). In general, the probability density function (PDF) of the
hygroscopicity parameter (κ) for 40–200 nm particles had a unimodal
distribution, and mean κ-PDF patterns for different sizes were
similar, suggesting that the particles were highly aged and internally mixed
because of strong photochemical reactions. The κ calculated from the
hygroscopic growth factor in the daytime and at night suggests that
photochemical reactions largely enhanced the aerosol hygroscopicity. This
effect became weaker as the particle size increased. In addition, the aerosol
hygroscopicity was much larger at XT than at other sites in the NCP. This is
because new particle formation takes place much more frequently in the
central NCP, which is heavily polluted from industrial activities, than
elsewhere in the region. The evolution of the planetary boundary layer played
a dominant role in dictating aerosol mass concentration. Particle size was
the most important factor influencing the ability of aerosols to activate,
whereas the effect of chemical composition was secondary, especially when
supersaturation was high. Using a fixed value of κ=0.31 to calculate
the cloud condensation nuclei number concentration in this region suffices.
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