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

A real quaternion spherical ensemble of random matrices

One can identify a tripartite classification of random matrix ensembles into geometrical universality classes corresponding to the plane, the sphere and the anti-sphere. The plane is identified with Ginibre-type (iid) matrices and the anti-sphere with truncations of unitary matrices. This paper focusses on an ensemble corresponding to the sphere: matrices of the form \bY= \bA^{-1} \bB, where \bA and \bB are independent $N\times N$ matrices with iid standard Gaussian real quaternion entries. By applying techniques similar to those used for the analogous complex and real spherical ensembles, the eigenvalue jpdf and correlation functions are calculated. This completes the exploration of spherical matrices using the traditional Dyson indices $\beta=1,2,4$. We find that the eigenvalue density (after stereographic projection onto the sphere) has a depletion of eigenvalues along a ring corresponding to the real axis, with reflective symmetry about this ring. However, in the limit of large matrix dimension, this eigenvalue density approaches that of the corresponding complex ensemble, a density which is uniform on the sphere. This result is in keeping with the spherical law (analogous to the circular law for iid matrices), which states that for matrices having the spherical structure \bY= \bA^{-1} \bB, where \bA and \bB are independent, iid matrices the (stereographically projected) eigenvalue density tends to uniformity on the sphere.Comment: 25 pages, 3 figures. Added another citation in version

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

A method to calculate correlation functions for β=1\beta=1 random matrices of odd size

The calculation of correlation functions for $\beta=1$ random matrix ensembles, which can be carried out using Pfaffians, has the peculiar feature of requiring a separate calculation depending on the parity of the matrix size N. This same complication is present in the calculation of the correlations for the Ginibre Orthogonal Ensemble of real Gaussian matrices. In fact the methods used to compute the $\beta=1$, N odd, correlations break down in the case of N odd real Ginibre matrices, necessitating a new approach to both problems. The new approach taken in this work is to deduce the $\beta=1$, N odd correlations as limiting cases of their N even counterparts, when one of the particles is removed towards infinity. This method is shown to yield the correlations for N odd real Gaussian matrices.Comment: 20 pages, corrected typo

Two-sided Grassmann-Rayleigh quotient iteration

The two-sided Rayleigh quotient iteration proposed by Ostrowski computes a pair of corresponding left-right eigenvectors of a matrix $C$. We propose a Grassmannian version of this iteration, i.e., its iterates are pairs of $p$-dimensional subspaces instead of one-dimensional subspaces in the classical case. The new iteration generically converges locally cubically to the pairs of left-right $p$-dimensional invariant subspaces of $C$. Moreover, Grassmannian versions of the Rayleigh quotient iteration are given for the generalized Hermitian eigenproblem, the Hamiltonian eigenproblem and the skew-Hamiltonian eigenproblem.Comment: The text is identical to a manuscript that was submitted for publication on 19 April 200

Intrinsic Statistics on Riemannian Manifolds: Basic Tools for Geometric Measurements

A preliminary appeared as INRIA RR-5093, January 2004.International audienceIn medical image analysis and high level computer vision, there is an intensive use of geometric features like orientations, lines, and geometric transformations ranging from simple ones (orientations, lines, rigid body or affine transformations, etc.) to very complex ones like curves, surfaces, or general diffeomorphic transformations. The measurement of such geometric primitives is generally noisy in real applications and we need to use statistics either to reduce the uncertainty (estimation), to compare observations, or to test hypotheses. Unfortunately, even simple geometric primitives often belong to manifolds that are not vector spaces. In previous works [1, 2], we investigated invariance requirements to build some statistical tools on transformation groups and homogeneous manifolds that avoids paradoxes. In this paper, we consider finite dimensional manifolds with a Riemannian metric as the basic structure. Based on this metric, we develop the notions of mean value and covariance matrix of a random element, normal law, Mahalanobis distance and X² law. We provide a new proof of the characterization of Riemannian centers of mass and an original gradient descent algorithm to efficiently compute them. The notion of Normal law we propose is based on the maximization of the entropy knowing the mean and covariance of the distribution. The resulting family of pdfs spans the whole range from uniform (on compact manifolds) to the point mass distribution. Moreover, we were able to provide tractable approximations (with their limits) for small variances which show that we can effectively implement and work with these definitions

Use of SMS texts for facilitating access to online alcohol interventions: a feasibility study

A41 Use of SMS texts for facilitating access to online alcohol interventions: a feasibility study In: Addiction Science & Clinical Practice 2017, 12(Suppl 1): A4

Structural basis for both pro- and anti-inflammatory response induced by mannose-specific legume lectin from Cymbosema roseum

Legume lectins, despite high sequence homology, express diverse biological activities that vary in potency and efficacy. In studies reported here, the mannose-specific lectin from Cymbosema roseum (CRLI), which binds N-glycoproteins, shows both pro-inflammatory effects when administered by local injection and anti-inflammatory effects when by systemic injection. Protein sequencing was obtained by Tandem Mass Spectrometry and the crystal structure was solved by X-ray crystallography using a Synchrotron radiation source. Molecular replacement and refinement were performed using CCP4 and the carbohydrate binding properties were described by affinity assays and computational docking. Biological assays were performed in order to evaluate the lectin edematogenic activity. The crystal structure of CRLI was established to a 1.8 Å resolution in order to determine a structural basis for these differing activities. The structure of CRLI is closely homologous to those of other legume lectins at the monomer level and assembles into tetramers as do many of its homologues. The CRLI carbohydrate binding site was predicted by docking with a specific inhibitory trisaccharide. CRLI possesses a hydrophobic pocket for the binding of α-aminobutyric acid and that pocket is occupied in this structure as are the binding sites for calcium and manganese cations characteristic of legume lectins. CRLI route-dependent effects for acute inflammation are related to its carbohydrate binding domain (due to inhibition caused by the presence of α-methyl-mannoside), and are based on comparative analysis with ConA crystal structure. This may be due to carbohydrate binding site design, which differs at Tyr12 and Glu205 position. © 2011 Elsevier Masson SAS. All rights reserved