An autophosphorylating but not transphosphorylating activity is associated with the unique N terminus of the herpes simplex virus type 1 ribonucleotide reductase large subunit.

We report on a protein kinase function encoded by the unique N terminus of the herpes simplex virus type 1 (HSV-1) ribonucleotide reductase large subunit (R1). R1 expressed in Escherichia coli exhibited autophosphorylation activity in a reaction which depended on the presence of the unique N terminus. When the N terminus was separately expressed in E. coli and partially purified, a similar autophosphorylation reaction was observed. Importantly, transphosphorylation of histones and of proteins in HSV-1-infected cell extracts was also observed with purified R1 and with truncated R1 mutants in which most of the N terminus was deleted. Ion-exchange chromatography was used to separate the autophosphorylating activity of the N terminus from the transphosphorylating activity of an E. coli contaminant protein kinase. We propose a putative function for this activity of the HSV-1 R1 N terminus during the immediate-early phase of virus replication.     

The laminated sediments of Loch Ness, Scotland: Preliminary report on the construction of a chronology of sedimentation and its potential use in assessing Holocene climatic variability

Two sediment cores, ca. 6 m long, have been recovered from the north basin of Loch Ness, Scotland. Each consists of ca. 4.5 m of laminated gyttja, terminating in a basal grey, unlaminated clay. A suite of three ¹⁴C AMS dates have been obtained, and place the base of the gyttja at ca. 9500 yr B.P. Investigations into the structure, composition and formation of the laminae are in progress. Analysis by Backscatter Scanning Electron Microscopy (BSEM) has demonstrated that they consist of couplets comprising dark, clay-rich sediments thought to be deposited from spring to autumn, and pale, silt-rich layers believed to represent sedimentation through winter. The laminae are thus thought to record incidence and intensity of streamflow into the Loch over the past nine millennia, and thus illustrate environmental change over the region for most of the Holocene. X-ray densitometry has been utilised in order to count the laminations and test the hypothesis that they are varves. It has, however, proved difficult to obtain a continuous sequence of countable laminations, although it has been possible to construct a fragmented, floating chronology which indicates that the hypothesis may be correct. Examination of lamination thickness reveals that although the average rate of sedimentation throughout the time periods studied seems to have remained fairly constant, significant variations have occurred. As Loch Ness is located on the northwest oceanic fringe of Europe, any climatic signal which the laminations contain will be closely related to even larger scale events over and within the North Atlantic Ocean, which is a major controller of global climate. Preliminary statistical investigation of sequences of laminae is being carried out in order to search for periodicity of sedimentation which may then be related to appropriate climatic indices.     

Identification and characterization of the cAMP binding proteins of yeast by photoaffinity labeling.

Modulation of a membrane glycoprotein, approximate molecular weight 200,000, in concert with active ionic flux has been shown in a human neuroblastoma cell line. The modulating agent was 2% dimethyl sulfoxide. Other neuronal properties, acetylcholinesterase and choline acetyltransferase, were also modulated but to a lesser extent. The appearance of this glycoprotein on the surface of both human and mouse neuroblastoma cells only under conditions of differentiation leads to the suggestion that it is directly involved with the active Na⁺ channels.     

Interaction of soluble pig heart glutamate-aspartate transaminase with various beta,gamma-unsaturated amino acids

beta-Ethylidene-DL-aspartate (beta EA) and beta-methylene-DL-glutamate (beta MG) were synthesized and tested as potential suicide inhibitors of soluble pig heart glutamate-aspartate transaminase (sGAT). beta MG was found to be a) a substrate with a very low turnover number relative to glutamate and b) a competitive inhibitor with respect to aspartate (albeit with a large binding constant). At high concentrations beta MG inactivated the enzyme but only very slowly. beta EA was also found to be a substrate with a very low turnover number; it did not inactivate the enzyme (1 hr, 25 degrees C) even at a high concentration. However, beta EA was found to bind to the enzyme with an affinity comparable to that of aspartate and glutamate. beta-Methylene-DL-aspartate (beta MA) has been shown to rapidly inactivate glutamate-aspartate transaminase. Therefore, it appears that glutamate-aspartate transaminase can bind analogues of aspartate with alkene groups in the beta position. The conjugated carbonyl groups of beta MA and beta EA will enhance Michael addition in comparison with that expected for vinylglycine. On the other hand, the presence of the methyl groups should reduce the electrophilicity of the double bond of beta EA compared to beta MA. This deactivation and/or steric hindrance to Michael attack may account for the inability of beta EA to inactivate sGAT. Therefore, it may be possible to design selective suicide inhibitors of glutamate-aspartate++ transaminase with the following structure: HO2CC(= CHX)CH(CO2H)NH2, where X is an electron-withdrawing group. Ideally, X would increase the reactivity of the double bond while affording a minimum of steric hindrance to susceptible enzyme-bound bases.