Classical Raman spectroscopic studies of NADH and NAD+ bound to lactate dehydrogenase by difference techniques

The binding of the coenzymes NAD+ and NADH to lactate dehydrogenase causes significant changes in the Raman spectra of both of these molecules relative to spectra obtained in the absence of enzyme. The molecular motions of the bound adenine moiety of both NAD+ and NADH as well as adenine containing analogues of these coenzymes produce Raman bands that are essentially identical, suggesting that the binding of adenine to the enzyme is the same regardless of the nicotinamide head-group nature. We also have observed that the molecular motions of the bound adenine moiety are different from both those obtained when it is in either water, various hydrophobic solvents, or various other solvent compositions. Protonation of the bound adenine ring at the 3-position is offered as a possible explanation. Significant shifts are observed in both the stretching frequency of the carboxamide carbonyl of NAD+ and the rocking motion of the carboxamide NH2 group of NADH. These shifts are probably caused by hydrogen bonding with the enzyme. The interaction energies of these hydrogen-bonding patterns are discussed. The aromatic nature of the nicotinamide moiety of NAD+ appears to be unchanged upon binding. Pronounced changes in the Raman spectrum of the nicotinamide moiety of NADH are observed upon binding; some of these changes are understood and discussed. Finally, these results are compared to analogous results that were recently reported for liver alcohol dehydrogenase [Chen et al. (1987) Biochemistry 26, 4776-4784]. In general, the coenzyme binding properties are found to be quite similar, but not identical, for the two enzymes.     

The specific binding activities of human urinary radioiodinated colony-stimulating factor-1 to various human tissue cells

1. Colony-stimulating factor (CSF-1) was isolated from a large volume of fresh normal human urine by 5 steps of purification and enrichment. 2. The purification factor is 100,000 fold and the purified compound exhibits a 2.16 x 10(7) U/mg of protein sp. act. 3. The isolated CSF-1 is a sialoglycoprotein with 41.5% of carbohydrate. The almost complete removal of this carbohydrate moiety (up to 91%) was achieved by incubation with trifluoromethane sulfonic acid. 4. The deglycosylated CSF-1 (DG-CSF-1) possesses an apparent Mr 38,000 compared to native CSF-1 with an initial Mr 57,000 (Goa et al., 1988). 5. The features of the interaction of radio-iodinated [125I]CSF-1 with single cell suspensions from various human tissues (bone marrow, spleen, blood, peritoneal cavity, alveolar lavage, lymph node and thymus), were studied. 6. The binding activity of peritoneal macrophages was the highest among the cells examined and erythrocytes, thymus and blood granulocytes showed no CSF-1 binding. 7. On incubation with [125I]CSF-1 at 0 degrees C, cellular binding of [125I]CSF-1 reached a stable maximum within 16 hr. This is in contrast to the association behaviour at higher temperature. 8. At 37 degrees C, cellular associated [125I]CSF-1 levels reached, within 90 min, an unstable maximum which was up to 10 times less than that occurring under the same conditions at 0 degree C. From the Scatchard plot analysis, we obtained the affinity constant and the number of receptor(s). 9. The binding site is sensitive to trypsin. 10. The receptor alone, (labelled by cross-linking to [125I]CSF-1 with di-succinylimidyl-suberate), is a polypeptide with an approx. Mr 110,000. 11. Our results showed that the receptor of CSF-1 is a tyrosin-kinase.     

香薷中的化学成分

从香[Eltholtzia ciliata(Thund.)Hyland]中分出14个化合物,用波谱和化学等方法确定为6-甲基三十三烷(Ⅰ),13-环己基二十六烷(Ⅱ),β-谷甾醇(Ⅲ),棕榈酸(Ⅳ_a),亚油酸(Ⅳ_b),亚麻酸(Ⅳ_c),熊果酸(Ⅴ),5-羟基-6、7-二甲氧基黄酮(Ⅵ),5-羟基-7、8-二甲氧基黄酮(Ⅶ),5、7-二羟基-4′-甲氧基黄酮(Ⅷ),5-羟基-7、4′-二甲氧基双氢黄酮醇(Ⅸ),β-谷甾醇-3-β-D-葡萄糖甙(Ⅹ),5-羟基-6-甲基-7-O-α-D-半乳吡喃糖双氢黄酮甙(Ⅺ),刺槐素-7-O-β-D-葡萄糖甙(Ⅻ)。其中化合物Ⅺ为新化合物,除(Ⅳ_b)和(Ⅳ_c)外,其余为首次从该属植物中得到。     

Flavonoids as superoxide scavengers and antioxidants

The superoxide anions scavenging activity and antioxidation of seven flavonoids--quercetin, rutin, morin, acacetin, hispidulin, hesperidin, and naringin--were studied. The superoxide anions were generated in a phenazin methosulphate-NADH system and were assayed by reduction of nitroblue tetrazolium. The scavenging activity ranked: rutin was the strongest, and quercetin and naringin the second, while morin and hispidulin were very weak. The concentration values yielding 50% inhibition of lipid peroxidation in mouse liver homogenate were in order of 10(-6) M for quercetin, rutin, and morin; and of 10(-5) M for acacetin and hispidulin, while naringin and hesperidin had no antioxidative action. In comparison with the antioxidative and scavenging activities of flavonoids, there are no correlations.     

金线鲃属鱼类一新种(鲤形目:鲤科)

作者于1986年6月在贵州盘县一溶洞的水体中采到金线鲃属鱼类一新种,命名为角金线鲃,新种Sinocyclocheilus angularis sp. nov.。本新种形状怪异,头背缘末端具一朝前的角状突,其内有顶骨突;眼正常;侧线鳞明显;鳔后室发达等特征而易与金线鲃属其他种相区别。     

紫茎泽兰的光合作用特征及其生态学意义

本文研究了紫茎泽兰(Eupatorium adenophorum Spreng)光合作用强度的变化规律及其与环境主要生态因子的关系,比较了它与某些农作物叶片净光合速率的差异,得到如下结果: 1.紫茎泽兰是一种阳性偏阴的C_3类草本植物,其光合作用的光饱和点约为40000 lx,光补偿点约为700 lx,且具有80 ppm左右的CO_2补偿点。 2.紫茎泽兰的最大净光合速率能达到23毫克CO_2/平方分米·时左右,叶片净光合速率的日变化规律呈双峰曲线型(主峰在10时左右,次峰在16时左右)。在一年中有较长的时间,它的光合速率保持着较高的水平。 3.生长在一般菜园土上的紫茎泽兰,当土壤含水量降至17％左右时,叶片光合速率接近0:而且,受过干旱处理的紫茎泽兰植株,在恢复供水后的第三天,其光合速率只达到原来的53％。 根据以上结果,结合受紫茎泽兰危害地区干湿季分明的特点,提出干季是防除紫茎泽兰的最佳季节。     

云南哀牢山南部鸟类的区系分析

哀牢山南延部分在云南省南部至东南部红河哈尼族彝族自治州(简称红河州)内,呈西北东南走向。南部为热带气候,北部为亚热带气候区。地势起伏虽不若滇西北三大峡谷之剧烈,但河口县与金平县内西隆山峰之高差却约达3,000米。这一南延部分区域内,植被类型复杂,花果繁茂,昆虫种类丰富。如此优越的自然条件,也导致了鸟类种类繁多昌盛的结果。     

白腹锦鸡鸣声的声谱分析

1986年1月—8月,12月及翌年1月,我们在云南省昆明市西部山区进行白腹锦鸡野外生态观察期间,录制了白腹锦鸡的鸣声。本文就啼叫声、呼唤声、惊叫声、恐惧叫声、威胁叫声及召唤雏鸟声等6种意义比较明确的鸣声进行了声谱分析,探讨各种鸣声与其相应的行为关系。