Properties of a New Plant Serine Protease Cucumisin

Cucumisin [EC 3.4.21.25] was coupled to cyanogen bromide-activated Sephayose 4B. The specific activity of the immobilized cucumisin was 41% of that of the soluble cucumisin toward casein. The immobilized enzyme was more stable against alkaline inactivation or heat than the soluble enzyme. In using affinity chromatography on a column of the immobilized cucumisin-Sepharose, cucumisin inhibitor was not obtained from potent sources of proteinase inhibitors, such as pig kidney and liver, avian and turtle egg-whites, or soybeans.     

Effect of streptolysin S on liposomes Influence of membrane lipid composition on toxin action

The effect of the bacterial cytolytic toxin, streptolysin S, on liposomes composed of various phospholipids was investigated. Large unilamellar vesicles containing [¹⁴C]sucrose were prepared by reverse-phase evaporation, and membrane damage produced by the toxin was measured by following the release of labeled marker. The net charge of the liposomes had little or no effect on their susceptibility to steptolysin S and the toxin was about equally effective on liposomes composed of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and phosphatidylglycerol. Experiments with liposomes composed of synthetic phospholipids showed that the ability of the toxin to produce membrane damage depended on the degree of unsaturation of the fatty acyl chains. The order of sensitivity was C18 : 2 phosphatidylcholine > C18 : 1 phosphatidylcholine > C18 : 0 phosphatidylcholine = C16 : 0 phosphatidylcholine. Liposomes containing the latter two phospholipids were virtually unaffected by streptolysin S, and experiments with C18 : 0 phosphatidylcholine suggested that toxin activity does not bind to liposomes composed of phospholipids with saturated fatty acyl chains. The inclusion of 40 mol% cholesterol in C16 : 0 phosphatidylcholine and C18 : 0 phosphatidylcholine liposomes made these vesicles sensitive to streptolysin S. Egg phosphatidylcholine liposomes, which were unaffected at 0°C and 4°C became susceptible to the toxin at these temperatures when cholesterol was included. Liposomes composed of C14 : 0 phosphatidylcholine were unaffected by streptolysin S at temperatures below the chain-melting transition temperature (23°C) of this phospholipid, but became increasingly susceptible above this temperature. The results suggest that the fluidity of the phospholipid hydrocarbon chains in the membrane is important in streptolysin S action.     

The ability of nonspecific T-cell stimulators to induce helper-cell-dependent increases in either polyclonal or isotype-restricted Ig production in vivo

In order to delineate the various roles of T cells in B-cell activation, mice were exposed to a variety of specific or nonspecific T-cell stimuli including mitogens, e.g., concanavalin A, adjuvants, e.g., complete Freund's adjuvant, and colchicine plus nonimmunogenic doses of antigen, anti-lymphocyte serum, and pathogens and their spleens analyzed for total class-specific immunoglobulin-secreting cells as indicators of helper cell generation. The results demonstrate that, depending on the mode of stimulation, markedly different Ig-secreting cell response patterns were induced, differing with respect to their kinetics and the isotype induced. In contrast to polyclonal T-cell stimuli such as concanavalin A and 17X lethal malaria which induced increases in all classes of Ig-recruiting cells, injection of many T-cell-activating agents resulted in the selective production of IgG clones in particular IgG 1. Such findings are discussed in terms of the different mechanisms of T-cell help and provide further evidence for functional heterogeneity in the T-helper-cell pool.     

Inhibition of co-operative enzymes by substrate-analogues: Possible implications for the physiological significance of negative co-operativity illustrated by phenylalanine metabolism in higher plants

The “Hill” equation for co-operative binding-systems has been extended to describe the effect of substrate-analogue on the binding of substrate to an oligomeric protein. It is demonstrated that the more negatively co-operative the binding-system, the more sensitive is the binding of substrate to inhibition by increases in the relative concentration of substrate-analogue. It is proposed that the physiological significance of negative co-operativity for enzymes may be complementary to the physiological significance of positive co-operativity. The effect of negative co-operativity is to make substrate binding more sensitive to inhibition by relative increases in the concentration of substrate-analogue (e.g. for many enzymes product of the reaction) at the expense of decreased sensitivity of substrate binding to relative changes in substrate concentration compared to a system with equivalent, independent substrate binding sites. In contrast, the effect of positive co-operativity is to make the enzyme more sensitive to relative changes in substrate concentration at the expense of decreased sensitivity to inhibition by relative increases in product concentration, compared to an enzyme without co-operative binding.     

The ichthyoplankton of a seasonally closed estuary in temperate Australia. Does an extended period of opening influence species composition?

Fish eggs and larvae were collected monthly between September 1987 and April 1989 from sites throughout the main basin and within the saline regions of the two main tributary rivers of Wilson Inlet, a seasonally closed estuary in south-western Australia. Of the eggs, 43.7% belonged to Engraulis australis (Shaw) and the rest to unidentified teleosts. The larval fish assemblage comprised 17 families represented by 25 species. The Gobiidae contained the highest number of species (four) and contributed approximately 57% of all larvae caught. Pseudogobius olorum (Sauvage) and E. australis were the most abundant species, contributing 43.9 and 27.9% to the total larval catch, respectively. The larvae of species which breed within Wilson Inlet dominated the assemblage, both in terms of number of species (64%) and contribution to total catch (99.9%). The numbers of the eight marine species and one freshwater species represented in the ichthyoplankton were very low. Classification and multi-dimensional scaling ordination showed that the composition of the larval fish fauna at the various sites during a period when the estuary remained open to the sea (December 1988-April 1989) was similar to that of the corresponding sites during the same period in the previous year when the estuary had become closed (December 1987-April 1988). This can be attributed to the spatial distribution, time of occurrence and abundance of estuarine-spawned larvae being similar in the two periods and to the rarity of marine-spawned larvae, even in the spring and summer of 1988/1989 when the estuary was open for the whole time when most marine teleosts spawn in south-western Australia. The low occurrence of marine-spawned larvae in Wilson Inlet reflects the fact that tidal water movement within the basin of the system is so small that it is unable to facilitate the transport and dispersion of larvae. The ichthyoplankton of Wilson Inlet resembles that of other poorly-flushed estuaries in that it is low in species richness and dominated by estuarine-spawned larvae.     

7-O-Galloyl-(+)-catechin and 3-O-galloylprocyanidin B-3 from Sanguisorba officinalis

7-O-Galloyl-(+)-catechin and 3-O-galloylprocyanidin B-3, along with gambiriins A-1 and B-3 and four polygalloylglucoses, have been isolated fro     

QTL identification of grain protein concentration and its genetic correlation with starch concentration and grain weight using two populations in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.)

Protein is one of the three main storage chemical components in maize grains, and is negatively correlated with starch concentration (SC). Our objective was to analyse the influence of genetic backgrounds on QTL detection for protein concentration (PC) and to reveal the molecular genetic associations between PC and both SC and grain weight (GWP). Two hundred and eighty-four (Pop1) and 265 (Pop2) F_2:3 families were developed from two crosses between one high-oil maize inbred GY220 and two normal maize inbreds 8984 and 8622 respectively, and were genotyped with 185 and 173 pairs of SSR markers. PC, SC and GWP were evaluated under two environments. Composite interval mapping (CIM) and multiple interval mapping (MIM) methods were used to detect single-trait QTL for PC, and multiple-trait QTL for PC with both SC and GWP. No common QTL were shared between the two populations for their four and one PC QTL. Common QTL with opposite signs of effects for PC and SC/GWP were detected on three marker intervals at bins 6.07–6.08, 8.03 and 8.03–8.04. Multiple-traits QTL mapping showed that tightly-linked QTL, pleiotropic QTL and QTL having effects with opposite directions for PC and SC/GWP were all observed in Pop1, while all QTL reflected opposite effects in Pop2.