C1q,a collagen-like complement subcomponent,in dermatosparactic cattle: its extracellular modification is not affected by lack of procollagen N-terminal proteinase (pN-proteinase)

C1q, a collagen-like complement protein, was purified from the serum of a ddermatosparactic calf which lacks procollagen N-terminal proteinase (pN-proteinase). The specific hemolytic activity of the serum Clq from the dermatosparactic animal was identical to that of C1q from a normal calf. Gel-filtration of serum from dermatosparactic calf, on Sepharose 6B, showed the presence of C1q-antigenic material at only one position which was identical to the elution position of normal bovine C1q. No differdence, under dissociating conditions, could be seen in the size of the chains of C1q in specific immunoprecipitates isolated from the sera of dermatosparactic and normal animals, as judged by polyacrylamidegel electrophoresis (PAGE) in the presence of sodium dodecyl sulfate (SDS). The C1q from the dermatosparactic animal showed the same N-terminal amino acid and typtic-digest peptide pattern on HPLC as C1q from the normal calf. These results strongly suggest that pN-proteinase is not involved in the extracellular processing of C1q.     

Effect of salinity on phosphate accumulation and injury in soybean

Many soybean [Glycine max (L.) Merr.] genotypes that are grown in solution cultures are highly sensitive to the combination of both salinity and inorganic phosphate (Pi) in the substrate. This effect has been observed on numerous occasions on plants grown in a saline medium that contained a substantial amount of Ca (i.e., CaCl₂/NaCl=0.5 on a molar basis). Because Ca is important in regulating ion transport and membrane permeability, solution culture experiments were designed to examine the effects of various concentrations of Pi and ratios of CaCl₂/NaCl (0 to 0.5 on a molar basis) at a constant osmotic potential (−0.34 MPa) on this adverse interaction. Four soybean cultivars (‘Lee’, ‘Lee 74’ ‘Clark’ and ‘Clark 63’) were tested. No adverse salinity x Pi interaction was found on Lee at any ratio and leaf P and Cl were maintained below 300 and 200 mmol kg⁻¹ dry wt, respectively. Clark, Clark 63 and Lee 74 soybean plants, on the other hand, were severely injured by solution salinity (−0.34 MPa osmotic potential) when substrate Pi was ≥0.12 mM. Reduced substrate Ca did not intensify the salinity x Pi interaction. On the contrary, the onset of injury was hastened and more severe with increased CaCl₂/NaCl ratios in isotonic solutions. Shoot and root growth rates decreased as injury increased. Leaf P concentrations from these cultivars grown in saline solutions with 0.12 mM Pi were excessive (>600 mmol kg⁻¹ dry wt) compared with concentrations commonly found in soybean leaf tissue yet they were independent of the severity of injury. Since leaf Cl increased wiht increased CaCl₂/NaCl ratio, we suspect that the severity of foliar injury was related to the combined effects of excessive P and Cl within the tissue. Lee 74, the only injured cultivar examined that excluded Cl from its leaves, was less sensitive than either Clark cultivar and its injury was characteristically different. Other ion interactions were reported that may have played a role in injury susceptibility.     

Transforming activities of sodium fluoride in cultured Syrian hamster embryo and BALB/3T3 cells

The transforming activity of sodium fluoride was studied in the SHE and the BALBl3T3 cell culture systems. Initiating and promoting activities were then investigated by means of the orthogonal methodology. Sodium fluoride was found to induce morphological transformation of SHE cells seeded on a feeder layer of X-irradiated cells at high concentrations (75–125 g/ ml). When the cells were seeded in the absence of a feeder-layer, the transformation frequencies increased in a dose-dependent manner with the concentrations of sodium fluoride ranging from 0 to the highly toxic concentration of 200 g/ml. In the BALBl3T3 cell system, sodium fluoride was negative in the standard Kakunaga procedure, while through the experiment designed by table L₈ (2⁷) of the orthogonal method, an initiating-like effect and a weak promoting activity were detected within the concentrations ranging from a 25 g/ ml to a 50 g/ ml concentration which is highly toxic for BALBl3T3 cells. From these results, it is suggested that, besides a genetic mode of action, sodium fluoride could possibly act through a non-genotoxic mechanism.Abbreviations CE cloning efficiency - NaF sodium fluoride - SHE Syrian hamster embryo - TF transformation frequency     

Effects of Salinity on the Extensibility and Ca Availability in the Expanding Region of Growing Barley Leaves

The growth of barley (Hordeum vulgare L.) leaves is reduced by salinity. We used the Instron extensometric technique to measure the reversible and irreversible compliance of the expanding regions of growing barley leaves from plants exposed to 1, 40, 80 and 120 mM NaCl in nutrient solution. Two barley cultivars differing in salinity resistance (cv ‘Arivat’ and cv ‘Briggs’) were compared over 5d of leaf growth. During the period of most active leaf expansion, salinity reduced reversible compliance and increased compliance in the leaf segments, although responses to salinity were complex and changed over the course of leaf expansion. Salinity increased irreversible compliance more in the salt-sensitive cultivar Arivat than in the more salt-tolerant cultivar Briggs. Elemental analysis of the basal leaf segments used for extensometry revealed an accumulation of Na and a depletion of Ca in segments from salinized plants, resulting in very high Na: Ca ratios in salinized expanding tissue. The concentrations of K and Mg in basal leaf tissue were elevated by salinity. Our data do support the hypothesis that the inhibition of leaf expansion by salinity stress is mediated by a decline in irreversible extensibility. We suggest that reduced Ca availability in expanding leaf tissue may contribute to growth reduction in salt-stressed barley seedlings.     

The principal islet of the coho salmon (Oncorhyncus kisutch) contains the BB isoenzyme of creatine kinase

The importance of creatine kinase (E.C. 2.7.3.2) in endocrine tissues has been generally overlooked. Using a specific radiometric assay, we have demonstrated the existence of CK in the Brockmann body (principal islet) of the Coho salmon. We have purified this protein from insular tissue and concurrently purified CK from brain and muscle of the salmon. Purification characteristics, immunological cross-reactivity, and N-terminal sequence analysis have demonstrated that the predominant cytosolic CK from the Brockmann body is indistinguishable from the BB (brain) isoenzyme. Immunocytochemical studies indicated that the enzyme resides in the endocrine parenchyma. Phosphocreatine may serve as a reservoir of energy in the islet and augment its capacity to secrete hormones. The induction of CK-BB in the islet by other hormones could influence the secretion of insular hormones. Interorgan flux of the substrate creatine may be an undescribed mechanism of physiological regulation.     

Stress studies in lentil (Lens esculenta Moench)

The response of soil exchangeable sodium percentage levels to nitrate reductase activity, nitrite reductase activity, free proline, DNA, RNA, chlorophyll a and b contents and yield components in lentil (Lens esculenta Moench)cv. PL 406 was studied in a replicated pot experiment. All the biochemical observations were recorded at four growth stages i.e. 30, 60, 90 and 120 days after sowing (DAS). Germination occurred up to exhangeable sodium percentage of 30, but plants survived only up to 25. With increasing exchangeable sodium percentage, there was a continuous decrease in chlorophyll a and b content, nitrate and nitrite reductase enzyme activities and DNA and RNA content. Increasing level of sodicity enhanced the free proline content up to 60 DAS, after which values fell.Number of pods per plant, 1000 grain weight and grain yield were significantly reduced with increasing level of sodicity, but the number of grains per pod was not affected.     

Hemoglobin Dallas (α97(G4)Asn→Lys):functional characterization of a high oxygen affinity natural mutant

Hemoglobin Dallas, an α-chain variant with a substitution of lysine for asparagine at position 97(G4), was found to have increased oxygen affinity ($\text{p}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{= 1}\text{mmHg at pH}\text{7.3}\text{and}\text{20°C), diminished cooperativity (0n,}\text{the Hill coefficient}\text{= 1.7}$) and reduced Bohr effect (about 50%). Addition of allosteric effectors (such as 2,3-diphosphoglycerate, inositol hexakisphosphate and bezafibrate) led to a decrease in oxygen affinity and increase in cooperative energy. Kinetic studies at pH 7.0 and 20°C revealed that (i), the overall rate of oxygen dissociation is 1.4-fold slower than that for HbA and (ii), the carbon monoxide dissociation rate is unaffected. The abnormal properties of this hemoglobin variant can be atttributed to a more ‘relaxed’ T-state.     

Calpain II induced insolubilization of lens β-crystallin polypeptides may induce cataract

Addition of calpain II (EC 3.4.22.17) to soluble proteins from 10-day-old rat lens caused an increase in turbidity and production of water-insoluble protein. The insolubilization increased with higher concentrations of both lens protein and calpain II, it could be prevented by the cysteine protease inhibitor E-64; it required at least 0.5 mM Ca²⁺, it was limited to 6% of the soluble protein present and resulted from precipitation β-crystallin polypeptides. When compared by two-dimensional electrophoresis, the insoluble β-crystallin polypeptides produced by calpain II were similar to insoluble β-crystallin polypeptides found incataractous lenses. Trypsin also caused insolubilization of β-crystallin polypeptides, but these polypeptides were unlike polypeptides produced during cataract formation. These data suggested that the loss of solubility was due to a specific removal of N/or C-terminal extensions from β-crystallin polypeptides by calpain II, and that a similar process may occur in vivo during cataract formation. It is hypothesized that the insoluble protein produced by calpain II causes cataract by increasing light scatter in the lens.     

表面活性剂的疏水性及电性对肌酸激酶的活力及复性能力的影响

测定了烷基硫酸钠(CnH_(2n+1)SO_4Na;记为CnS;n=8,10,12)和溴化十烷基三甲铵C_(10)H_(21)(CH_3)Br;记为C_(10)NM_3)对肌酸激酶(CreatineKinase;记为C.K.)的活力,以及在它们中变性后复活能力的影响。CnS对C.K.的变性效率随n的增加而增加,变性效率的对数和n之间有线性关系;CnS水C.K.的变性能力远大于C_(10)NM_3;C.K.被C_(10)NM_3变性以后,其复性能力(稀释时恢复活力的程度远大于C.K.被CnS变性后活力的恢复能力。这种差别主要是由于C_(10)NM_3带正电,而CnS带负电引起的。     

人肌肌酸激酶胍变性时的失活与构象变化的比较研究

应用二阶导数光谱、紫外差吸收光谱和荧光光谱等监测手段,研究了人肌肌酸激酶在盐酸胍溶液中的构象变化。二阶导数光谱结果表明,若以6M盐酸胍中肌酸激酶酪氨酸残基的暴露程度为100％,则天然酶酪氨酸残基的暴露程度只有2％。而紫外差吸收光谱和荧光光谱的变化与兔肌肌酸激酶的结果相似。比较不同胍浓度下人肌肌酸激酶的失活与构象变化,表明酶的失活先于构象变化。同时还测定了不同浓度胍溶液中人肌酶的失活与构象变化的速度常数。结果表明以几种方法测定的构象变化均为单相的一级过程,而酶的失活却呈现了由快慢两相组成的一级反应过程。比较同浓度胍溶液中的失活速度与构象变化速度,发现酶失活的快相反应速度常数比构象变化的速度常数大1—2个数量级,慢相速度常数与构象变化速度常数相近。上述结果进一步支持了酶的活性部位构象柔性的观点。