A guanosine 5′-triphosphate-dependent protein kinase is localized in the outer envelope membrane of pea chloroplasts

A guanosine 5-triphosphate (GTP)-dependent protein kinase was detected in preparations of outer chloroplast envelope membranes of pea (Pisum sativum L.) chloroplasts. The protein-kinase activity was capable of phosphorylating several envelope-membrane proteins. The major phosphorylated products were 23- and 32.5-kilo-dalton proteins of the outer envelope membrane. Several other envelope proteins were labeled to a lesser extent. Following acid hydrolysis of the labeled proteins, most of the label was detected as phosphoserine with only minor amounts detected as phosphothreonine. Several criteria were used to distinguish the GTP-dependent protein kinase from an ATP-dependent kinase also present in the outer envelope membrane. The ATP-dependent kinase phosphorylated a very different set of envelope-membrane proteins. Heparin inhibited the GTP-dependent kinase but had little effect upon the ATP-dependent enzyme. The GTP-dependent enzyme accepted phosvitin as an external protein substrate whereas the ATP-dependent enzyme did not. The outer membrane of the chloroplast envelope also contained a phosphotransferase capable of transferring labeled phosphate from [-32P]GTP to ADP to yield (-32P]ATP. Consequently, addition of ADP to a GTP-dependent protein-kinase assay resulted in a switch in the pattern of labeled products from that seen with GTP to that typically seen with ATP

Adenylate effects on protein phosphorylation in the interenvelope lumen of pea chloroplasts

A 64-kilodalton (kDa) protein, situated in the lumen between the inner and outer envelopes of pea (Pisum sativum L.) chloroplasts (Soll and Bennett 1988, Eur. J. Biochem., 175, 301–307) is shown to undergo reversible phosphorylation in isolated mixed envelope vesicles. It is the most conspicuously labelled protein after incubation of envelopes with 33 nmol·1-1 [-32P]ATP whereas incubation with 50 mol·1-1 [-32P]ATP labels most prominently two outer envelope proteins (86 and 23 kDa). Half-maximum velocity for phosphorylation of the 64-kDa protein occurs with 200 nmol·1-1 ATP, and around 40 mol·1-1 ATP for phosphorylation of the 86- and 23-kDa proteins, indicating the operation of two distinct kinases. GGuanosine-, uridine-, cytidine 5-triphosphate and AMP are poor inhibitors of the labelling of the 64-kDa protein with [-32P]ATP. On the other hand, ADP has a potent influence on the extent of labelling (half-maximal inhibition at 1–5 mol·1-1). The ADP-dependent appearance of 32P in ATP indicates that ADP acts by reversal of kinase activity and not as a competitive inhibitor. However, the most rapid loss of 32P from pre-labelled 64-kDa protein occurs when envelope vesicles are incubated with ATP t1/2=15 s at 20 molsd1-1 ATP). This induced turnover of phosphate appears to be responsible for the rapid phosphoryl turnover seen in situ

Sexual reproduction of human fungal pathogens

We review here recent advances in our understanding of sexual reproduction in fungal pathogens that commonly infect humans, including Candida albicans, Cryptococcus neoformans/gattii, and Aspergillus fumigatus. Where appropriate or relevant, we introduce findings on other species associated with human infections. In particular, we focus on rapid advances involving genetic, genomic, and population genetic approaches that have reshaped our view of how fungal pathogens evolve. Rather than being asexual, mitotic, and largely clonal, as was thought to be prevalent as recently as a decade ago, we now appreciate that the vast majority of pathogenic fungi have retained extant sexual, or parasexual, cycles. In some examples, sexual and parasexual unions of pathogenic fungi involve closely related individuals, generating diversity in the population but with more restricted recombination than expected from fertile, sexual, outcrossing and recombining populations. In other cases, species and isolates participate in global outcrossing populations with the capacity for considerable levels of gene flow. These findings illustrate general principles of eukaryotic pathogen emergence with relevance for other fungi, parasitic eukaryotic pathogens, and both unicellular and multicellular eukaryotic organisms

Collisional damping of spherical ice particles

This paper presents experimental values for the coefficient of restitution (en) for millimeter-sized ice particles colliding with massive walls at different temperatures. Three different wall materials are tested: hardened glass, ice and Acrylonitrile butadiene styrene (ABS) polymer. The results show a high sensitivity to impact velocity Vi, where en decreases rapidly with increasing Vi. The results also show a decrease in en with increasing temperature T. A novel model that predicts en based on the assumption of collisional melting and viscous damping caused by an increased premelted liquid-layer, is proposed. The model predicts both the velocity and the temperature trends seen in the experiments. The difference obtained in experiments between wall materials is also captured by the new model. A generalized regime map for ice particle collisions is proposed to combine the new model with previous work

Site of methylation of 2-phytyl-1,4-naphthoquinol in phylloquinone (vitamin K1) synthesis in spinach chloroplasts

Phylloquinol (the quinol form of vitamin K1) is synthesized from 2-phytyl-1,4-naphthoquinol and S-adenosylmethionine at the thylakoid membranes of spinach chloroplasts. The addition of soluble stroma protein (chloroplast extract) is necessary S-Adenosylhomocysteine acts as strong competitive inhibitor

Pair Plasma Dominance in the Parsec-Scale Relativistic Jet of 3C345

We investigate whether a pc-scale jet of 3C345 is dominated by a normal plasma or an electron-positron plasma. We present a general condition that a jet component becomes optically thick for synchrotron self-absorption, by extending the method originally developed by Reynolds et al. The general condition gives a lower limit of the electron number density, with the aid of the surface brightness condition, which enables us to compute the magnetic field density. Comparing the lower limit with another independent constraint for the electron density that is deduced from the kinetic luminosity, we can distinguish the matter content. We apply the procedure to the five components of 3C345 (C2, C3, C4, C5, and C7) of which angular diameters and radio fluxes at the peak frequencies were obtainable from literature. Evaluating the representative values of Doppler beaming factors by their equipartition values, we find that all the five components are likely dominated by an electron-positron plasma. The conclusion does not depend on the lower cutoff energy of the power-law distribution of radiating particles.Comment: 17 page

The protein import apparatus of chloroplasts

Routing of cytosolically synthesized precursor proteins into chloroplasts is a specific process which involves a multitude of soluble and membrane components. In this review we wil1 focus on early events of the translocation pathway of nuclear coded plastidic precursor proteins and compare import routes for polypeptide of the outer chloroplast envelope to that of internal chloroplast compartments. A number of proteins housed in the chloroplast envelopes have been implied to be involved in the translocation process, but so far a certain function has not been assigned to any of these proteins. The only exception could be an envelope localized hsc 70 homologue which could retain the import competence of a precursor protein in transit into the organelle