Chloroplasts require protein translocons at the outer and inner envelope membranes, termed TOC and TIC, respectively, to import thousands of cytoplasmically synthesized preproteins. However, the molecular identity of the TIC translocon remains controversial. Tic20 forms a 1-megadalton complex at the inner membrane and directly interacts with translocating preproteins. We purified the 1-megadalton complex from Arabidopsis, comprising Tic20 and three other essential components, one of which is encoded by the enigmatic open reading frame ycf1 in the chloroplast genome. All four components, together with well-known TOC components, were found stoichiometrically associated with different translocating preproteins. When reconstituted into planar lipid bilayers, the purified complex formed a preprotein-sensitive channel. Thus, this complex constitutes a general TIC translocon.

Site-directed mutants of cytochrome P-450cam (the cytochrome P-450 that acts as the terminal monooxygenase in the d-camphor monooxygenase system), in which threonine-252 had been changed to alanine, valine, or serine, were employed to study the role of the hydroxy amino acid in the monooxygenase reaction. The mutant enzymes were expressed in Escherichia coli and were purified by a conventional method. All the mutant enzymes in the presence of d-camphor exhibited optical absorption spectra almost indistinguishable from those of the wild-type enzyme in their ferric, ferrous, oxygenated, and carbon monoxide ferrous forms. In a reconstituted system with putidaredoxin and its reductase, the alanine enzyme consumed O2 at a rate (1100 per min per heme) comparable to that of the wild-type enzyme (1330 per min per heme), whereas the amount of exo-5-hydroxycamphor formed was less than 10% of that formed by the wild-type enzyme. About 85% of the O2 consumed was recovered as H2O2. The valine enzyme also exhibited an oxidase activity to yield H2O2 accompanied by a relative decrease in the monooxygenase activity. On the other hand, the serine enzyme exhibited essentially the same monooxygenase activity as that of the wild-type enzyme. Thus, uncoupling of O2 consumption from the monooxygenase function was produced by the substitution of an amino acid without a hydroxyl group. When binding of O2 to the ferrous forms was examined, the alanine and valine enzymes formed instantaneously an oxygenated form, which slowly decomposed to the ferric form with rates of 5.5 and 3.2 x 10(-3) sec-1 for the former and latter enzymes, respectively. Since these rates were too slow to account for the overall rates of O2 consumption, the formation of H2O2 was considered to proceed not by way of this route but through the decomposition of a peroxide complex formed by reduction of the oxygenated form by reduced putidaredoxin. Based on these findings, a possible mechanism for oxygen activation in this monooxygenase reaction has been discussed.

To cope with life‐threatening high osmolarity, yeast activates the high‐osmolarity glycerol (HOG) signaling pathway, whose core element is the Hog1 MAP kinase cascade. Activated Hog1 regulates the cell cycle, protein translation, and gene expression. Upstream of the HOG pathway are functionally redundant SLN1 and SHO1 signaling branches. However, neither the osmosensor nor the signal generator of the SHO1 branch has been clearly defined. Here, we show that the mucin‐like transmembrane proteins Hkr1 and Msb2 are the potential osmosensors for the SHO1 branch. Hyperactive forms of Hkr1 and Msb2 can activate the HOG pathway only in the presence of Sho1, whereas a hyperactive Sho1 mutant activates the HOG pathway in the absence of both Hkr1 and Msb2, indicating that Hkr1 and Msb2 are the most upstream elements known so far in the SHO1 branch. Hkr1 and Msb2 individually form a complex with Sho1, and, upon high external osmolarity stress, appear to induce Sho1 to generate an intracellular signal. Furthermore, Msb2, but not Hkr1, can also generate an intracellular signal in a Sho1‐independent manner.

Chloroplasts import thousands of nucleus-encoded preproteins synthesized in the cytosol through the TOC and TIC translocons on the outer and inner envelope membranes, respectively. Preprotein translocation across the inner membrane requires ATP; however, the import motor has remained unclear. Here, we report that a 2-MD heteromeric AAA-ATPase complex associates with the TIC complex and functions as the import motor, directly interacting with various translocating preproteins. This 2-MD complex consists of a protein encoded by the previously enigmatic chloroplast gene ycf2 and five related nuclear-encoded FtsH-like proteins, namely, FtsHi1, FtsHi2, FtsHi4, FtsHi5, and FtsH12. These components are each essential for plant viability and retain the AAA-type ATPase domain, but only FtsH12 contains the zinc binding active site generally conserved among FtsH-type metalloproteases. Furthermore, even the FtsH12 zinc binding site is dispensable for its essential function. Phylogenetic analyses suggest that all AAA-type members of the Ycf2/FtsHi complex including Ycf2 evolved from the chloroplast-encoded membrane-bound AAA-protease FtsH of the ancestral endosymbiont. The Ycf2/FtsHi complex also contains an NAD-malate dehydrogenase, a proposed key enzyme for ATP production in chloroplasts in darkness or in nonphotosynthetic plastids. These findings advance our understanding of this ATP-driven protein translocation system that is unique to the green lineage of photosynthetic eukaryotes.

We report the first evidence of a hot corino in a Bok globule. This is based on the ALMA observations in the 1.2 mm band toward the low-mass Class 0 protostar IRAS 19347+0727 in B335. Saturated complex organic molecules (COMs), CH 3 CHO, HCOOCH 3 , and NH 2 CHO, are detected in a compact region within a few 10 au around the protostar. Additionally, CH 3 OCH 3 , C 2 H 5 OH, C 2 H 5 CN, and CH 3 COCH 3 are tentatively detected. Carbon-chain related molecules, CCH and c-C 3 H 2 , are also found in this source, whose distributions are extended over a few 100 au scale. On the other hand, sulfur-bearing molecules CS, SO, and SO 2 , have both compact and extended components. Fractional abundances of the COMs relative to H 2 are found to be comparable to those in known hot-corino sources. Though the COMs lines are as broad as 5-8 km s −1 , they do not show obvious rotation motion in the present observation. Thus, the COMs mainly exist in a structure whose distribution is much smaller than the synthesized beam (0. 58 × 0. 52).

Context. Hot corinos are extremely rich in complex organic molecules (COMs). Accurate abundance measurements of COMs in such objects are crucial to constrain astrochemical models. In the particular case of close binary systems this can only be achieved through high angular resolution imaging. Aims. We aim to perform an interferometric study of multiple COMs in NGC1333 IRAS 4A, which is a protostellar binary hosting hot corino activity, at an angular resolution that is sufficient to distinguish easily the emission from the two cores separated by 1.8 . Methods. We used the Atacama Large (sub-)Millimeter Array (ALMA) in its 1.2 mm band and the IRAM Plateau de Bure Interferometer (PdBI) at 2.7 mm to image, with an angular resolution of 0.5 (120 au) and 1 (235 au), respectively, the emission from 11 different organic molecules in IRAS 4A. This allowed us to clearly disentangle A1 and A2, the two protostellar cores. For the first time, we were able to derive the column densities and fractional abundances simultaneously for the two objects, allowing us to analyse the chemical differences between them. Results. Molecular emission from organic molecules is concentrated exclusively in A2, while A1 appears completely devoid of COMs or even simpler organic molecules, such as HNCO, even though A1 is the strongest continuum emitter. The protostellar core A2 displays typical hot corino abundances and its deconvolved size is 70 au. In contrast, the upper limits we placed on COM abundances for A1 are extremely low, lying about one order of magnitude below prestellar values. The difference in the amount of COMs present in A1 and A2 ranges between one and two orders of magnitude. Our results suggest that the optical depth of dust emission at these wavelengths is unlikely to be sufficiently high to completely hide a hot corino in A1 similar in size to that in A2. Thus, the significant contrast in molecular richness found between the two sources is most probably real. We estimate that the size of a hypothetical hot corino in A1 should be less than 12 au. Conclusions. Our results favour a scenario in which the protostar in A2 is either more massive and/or subject to a higher accretion rate than A1, as a result of inhomogeneous fragmentation of the parental molecular clump. This naturally explains the smaller current envelope mass in A2 with respect to A1 along with its molecular richness. The extremely low abundances of organic molecules in A1 with respect to those in A2 demonstrate that the dense inner regions of a young protostellar core lacking hot corino activity may be poorer in COMs than the outer protostellar envelope.

To date, about two dozen low-mass embedded protostars exhibit rich spectra with lines of complex organic molecules (COMs). These protostars seem to possess a different enrichment in COMs. However, the statistics of COM abundance in low-mass protostars are limited by the scarcity of observations. This study introduces the Perseus ALMA Chemistry Survey (PEACHES), which aims at unbiasedly characterizing the chemistry of COMs toward the embedded (Class 0/I) protostars in the Perseus molecular cloud. Of the 50 embedded protostars surveyed, 58% of them have emission from COMs. 56%, 32%, and 40% of the protostars have CH 3 OH, CH 3 OCHO, and N-bearing COMs, respectively. The detectability of COMs depends neither on the averaged continuum brightness temperature, a proxy of the H 2 column density, nor on the bolometric luminosity and the bolometric temperature. For the protostars with detected COMs, CH 3 OH has a tight correlation with CH 3 CN, spanning more than two orders of magnitude in column densities normalized by the continuum brightness temperature, suggesting a chemical relation between CH 3 OH and CH 3 CN and a large chemical diversity in the PEACHES samples at the same time. A similar trend with more scatter is also found between all identified COMs, which hints at a common chemistry for the sources with COMs. The correlation between COMs is insensitive to the protostellar properties, such as the bolometric luminosity and the bolometric temperature. The abundance of larger COMs (CH 3 OCHO and CH 3 OCH 3 ) relative to that of smaller COMs (CH 3 OH and CH 3 CN) increases with the inferred gas column density, hinting at an efficient production of complex species in denser envelopes.

We report molecular line observations of the NGC 1333 IRAS 4C outflow in the Perseus Molecular Cloud with the Atacama Large Millimeter/Submillimeter Array. The CCH and CS emission reveal an outflow cavity structure with clear signatures of rotation with respect to the outflow axis. The rotation is detected from about 120 au up to about 1400 au above the envelope/disk mid-plane. As the distance to the central source increases, the rotation velocity of the outflow decreases while the outflow radius increases, which gives a flat specific angular momentum distribution along the outflow.The mean specific angular momentum of the outflow is about 100 au km s −1 . Based on reasonable assumptions on the outward velocity of the outflow and the protostar mass, we estimate the range of outflow launching radii to be 5−15 au. Such a launching radius rules out that this outflow is launched as an X-wind, but rather, it is more consistent arXiv:1808.00346v1 [astro-ph.GA] 1 Aug 2018 2 Zhang et al.to be a slow disk wind launched from relatively large radii on the disk. The radius of the centrifugal barrier is roughly estimated, and the role of the centrifugal barrier in the outflow launching is discussed.