Light-induced de Novo Synthesis of Ribulose 1,5-Diphosphate Carboxylase in Greening Leaves of Barley

An antibody specific for ribulose 1,5-diphosphate carboxylase was used to isolate the enzyme from greening barley (Hordeum vulgare L.) leaves. The increase in enzymatic activity during greening was due to de novo synthesis of the enzyme. Increases in enzymatic activity were accompanied by corresponding increases in enzyme protein and by incorporation of radioactive leucine, all of which were inhibited by low concentrations of cycloheximide. ¹⁴C-Labeled amino acids were incorporated into the enzyme by covalent peptide bonding.     

Diagnostic Accuracy in Venous Thrombosis

A group of 111 surgical patients at high risk of venous thrombosis were studied after operation by independent clinical assessment and with ¹²⁵I-fibrinogen to detect venous thrombosis. Almost half of the patients developed venous thrombosis. Of these, two-thirds were not suspected clinically despite careful scrutiny. In the patients in whom a clinical diagnosis of venous thrombosis was made this diagnosis was falsely positive in a quarter. More than half of all thrombotic episodes were detectable on the day after operation.The prevalence of venous thrombosis, together with the difficulty in diagnosing it, strongly supports the argument that a reduction in the incidence of pulmonary embolism must depend on widespread adoption of effective prophylaxis, especially in the large number of patients at high risk of venous thrombosis. Prophylactic trials must be objectively assessed, and it is in this field that the ¹²⁵I-fibrinogen technique probably has the most to offer.     

INFLUENCE OF AGE ON THE INCORPORATION OF [14C]GLYCINE INTO PROTEIN IN CHICKEN PERIPHERAL NERVE

Abstract—

• 1 The conditions for incorporation of [¹⁴C]glycine in vitro into proteins in the sciatic nerve of chickens have been studied and found to be similar to those of rat nerve.
• 2 Its incorporation decreases, however, linearly with age.
• 3 The content of RNA and of DNA of peripheral nerve and the RNA/DNA ratio alter linearly with age.
• 4 There is also a linear relationship between the specific radioactivity of the protein extract and the RNA content of the nerve.
• 5 There is a linear decline with age in the specific radioactivity of the protein fraction when expressed against the DNA content.
• 6 A linear relationship exists between the logarithm of the specific radioactivity and the length of the femur.

     

Kinetic studies on the interaction of alpha 1-proteinase inhibitor (Pittsburgh) with trypsin-like serine proteinases

The rates of interaction of a number of serine proteinases with a mutant form of alpha 1-proteinase inhibitor (referred to as alpha 1-proteinase inhibitor (Pittsburgh)), in which a methionine-358 to arginine-358 mutation has occurred, have been determined. An approximately 6,000-fold increase in the second order association rate constant with human thrombin was observed (48 M-1 X s-1 for the normal protein to 3.1 X 10(5) M-1 X s-1 for the arginine mutant), confirming previously observed data using bovine thrombin (Owen, M.C., Brennan, S.O., Lewis, J.H. & Carrell, R.W. (1983) New England J. Med. 309, 694-698). However, substantial increases in the rates of association with other trypsin-like enzymes were also noted, indicating that the replacement of methionine by a basic residue affects all serine proteinases with this kind of specificity. There was a marked decrease in the rates of interaction of the Pittsburgh mutant with both human neutrophil elastase and porcine pancreatic elastase, the inhibitor being converted into lower molecular mass fragments after interaction with either enzyme. Butanedione caused a substantial loss in the inhibitory activity of the arginine mutant, while having no effect on the normal protein. These data, when compared to those previously reported for differences in reaction rates between normal and oxidized alpha 1-proteinase inhibitor (Beatty, K., Bieth, J. & Travis, J. (1980) J. Biol. Chem. 255, 3931-3934), are consistent with the interpretation that the amino acid in the P1-position at the reactive site of this protein has a marked effect on determining its primary specificity.     

Glutamate receptor binding to cat central nervous system membranes

L-[3H]Glutamate exhibited specific binding to fresh membranes of cat CNS under physiological conditions of pH and temperature. This binding occurred in the absence of sodium ions. Kinetic analysis of the data for cerebellum suggested the presence of two distinct binding sites: a high-affinity process (Kd = 0.33 microM) with a capacity of 15 pmol/mg protein and a low-affinity process (Kd = 1.8 microM) which had a capacity of 65 pmol/mg protein. Several structural analogues of glutamic acid were able to appreciably inhibit the binding of [3H]glutamate. The distribution of glutamate binding between 12 regions of the CNS was measured. The amygdaloid complex exhibited the highest binding followed by hippocampus > hypothalamus identical to visual cortex identical to thalamus identical to caudate nucleus > olfactory bulb identical to tectum identical to cerebellum > dorsal pons identical to medulla > cervical spinal cord. These findings are consistent with the binding of [3H]glutamate being to its receptor.     

INFLUENCE OF AGE IN THE INCORPORATION OF [14C]GLYCINE INTO ISOLATED RAT NERVE SEGMENTS IN VITRO

—(1) Sciatic nerves of young rats have been shown to incorporate [¹⁴C]glycine in vitro into the protein fraction at a higher rate than nerves from adult rats under the same conditions. (2) Whilst there is little change in the DNA content of the sciatic nerve of rats with age, there is a is noticeable decline in the RNA content with age. (3) There a linear relationship between the specific activity of the protein fraction and the RNA content of the sciatic nerve under in vitro conditions. (4) There is a decline in the specific activity of the protein fraction with age when expressed against DNA. (5) A linear relationship exists between the logarithm of the specific activity and the length of the femur.     

Androgen dependence of specific kallikrein gene family members expressed in rat prostate

We have used oligonucleotide probes specific for members of the rat kallikrein/tonin gene family (PS, S1, S2, S3, K1, and P1) to establish which arginyl esteropeptidase (kallikrein-like) genes are expressed in the prostate. We have also compared the expression and androgen dependence of these genes in prostate, submaxillary gland (SMG) and kidney. Only S3 (tonin-like) and P1 (kallikrein-like) are expressed in the prostate, with S3 very much more abundant. Prostatic S3 mRNA disappears after 8 days castration and is restored to intact levels by dihydrotestosterone (DHT) but not estradiol benzoate (EB) for 8 days. Prostate P1 mRNA levels were similarly but not identically affected. All six genes are expressed in the SMG, with PS (true kallikrein) the most abundant. Levels of PS mRNA in SMG are unaffected by castration, DHT, or EB treatment, although mRNA levels of other kallikrein-like (S1, K1, and P1), tonin (S2), and tonin-like (S3) genes fall 40-60% after castration, and are unaffected or partially restored by DHT and/or EB administration. Only PS and K1 are expressed in the kidney, at much lower levels than in the SMG and unaffected by castration or steroids. These studies thus confirm and extend the concept of tissue specificity of arginyl esteropeptidase gene expression, and further demonstrate that the same gene(s) is differentially regulated by androgens in the rat prostate, SMG, and kidney.     

Kallikrein-gene expression in the rat gastrointestinal tract.

The serine proteinase glandular kallikrein has been demonstrated in the gastrointestinal tract, although there is some doubt as to whether it is synthesized there or derives from exocrine-gland secretions. Using a rat pancreatic kallikrein cRNA probe we have demonstrated kallikrein-like gene expression in the corpus, duodenum, jejunum, ileum, caecum and colon, and compared the pattern of expression with that of the gastrointestinal peptides somatostatin, gastrin and glucagon. In addition, using a panel of oligonucleotide probes specific for various members of the rat kallikrein-gene family, we have shown that the kallikrein-like gene expressed appears to be expressed as true kallikrein.     

A small basic ribosomal protein in Sulfolobus solfataricus equivalent to L46 in yeast: structure of the protein and its gene

The structure of the gene for a small, very basic ribosomal protein in Sulfolobus solfataricus has been determined and the structure of the protein coded by this gene (L46e) has been confirmed by partial amino acid sequencing. The protein shows substantial sequence homology to the eukaryotic ribosomal proteins L39 in rat and L46 in yeast. There is no sequence homology to any of the eubacterial ribosomal proteins suggesting that this protein is absent in the eubacterial ribosome.     

Optimising diquat use for submerged aquatic weed management

An experimentally derived prediction tool is under development which aims to assess potential deactivation of diquat caused by water and deposits on plant leaf surfaces in New Zealand water bodies, where aquatic weeds are targeted for diquat treatment. Optimising the use and success of diquat is important not only in managing public confidence in the use of aquatic herbicides, but also in minimising financial risk from failed treatments. Our approach focuses on characterising lake water quality and plant condition factors in these lakes to identify parameters that might be useful indicators of diquat deactivation potential. Water samples have been collected at 3-month intervals from lakes receiving large scale treatment for weed control. Samples have been analysed for turbidity, suspended solids, chlorophyll a, conductivity and dissolved anions. Samples have also been spiked with 1 mg l⁻¹ diquat to measure loss from adsorption and/or absorption. Shoot samples were also collected from targeted weed species at each sampling site and the amount of organic and inorganic deposits on plants has been measured and then added to a second diquat spiked sample to assess potential additional diquat loss from these deposits. Our results have shown deactivation from deposits on plant surfaces which is highly correlated with turbidity, including inorganic suspended solids and total suspended solids. A plant “dirtiness” scale has been devised to help predict the likely success or risk of diquat failure prior to any decision to proceed with treatment. Deactivation in water was only weakly linked to total suspended solids. Our failure to find significant correlation with the water quality factors measured may reflect the need for more detailed analysis of the particle size and composition of suspended solids and future research will address these issues.