Effect of different doses of chromium picolinate on protein metabolism in infant rats.

This 12-day study was conducted to evaluate the effects of three different levels of dietary chromium (100, 200, and 500 microg/day) in the form of chromium picolinate (CrPic) on growth and protein use in weaned rats. No significant effect of CrPic on body weight gain, food intake, or food conversion rate was observed. Elevated doses of CrPic seemed to increase muscle mass, either by stimulating protein anabolism by activation of insulin by chromium or by lowering protein degradation. However, these effects had no repercussions on overall growth, suggesting that any anabolic effect of chromium due to the action of insulin was probably marginal.     

氟烷对慢性缺氧兔心肌β肾上腺素能受体的影响

用放射配基结合法和高效液相色谱电化学法分别测定氟烷对慢性缺氧免心肌β肾上腺素能受体（简称β受体）和血浆儿茶酚胺的影响。结果表明：慢性缺氧后兔心肌β受体密度明显下降,受体亲和力无明显变化,血浆肾上腺素、去甲肾上腺素明显升高。缺氧吸入氟烷后兔心肌β受体密度进一步下降,亲和力增加,血浆肾上腺素、去甲肾上腺素无进一步变化。常氧吸入氟烷后心肌β受体密度无明显变化,亲和力升高,血浆肾上腺素、去甲肾上腺素反而降低。结果提示,氟烷抑制常氧兔交感神经活动而不能抑制缺氧兔交感神经活动,氟烷不改变常氧兔心肌β受体密度,但降低缺氧兔心肌β受体密度。β受体密度降低与缺氧后升高的儿茶酚胺下调β受体数目有关,同时可能与氟烷改变了缺氧心肌细胞膜脂质流动性使受体易向膜内移动有关。     

喜马拉雅灰包菇──相传是“喜马拉雅雪人”的一种食物

喜马拉雅灰包菇ＳｅｃｏｔｉｕｍｈｉｍａｌａｉｃｕｍＺａｎｇｅｔＤｏｉ,由Ｄｒ．ＴｅｉｚｏＯｇａｗａ采于尼泊尔境内的喜马拉雅山带,据当地居民云：该菌为喜马拉雅雪人的一种食物。所谓雪人可能是高山雪线一带的动物。本文对该菌的分类特征和雪人的有关讨论作了介绍。原模式存日本国立科学博物馆。     

Hydrophobic interaction chromatography of Micrococcus lysodeikticus F1-ATPase. A method to detect conformational flexibility of the molecule

Hydrophobic interaction chromatography of purified ATPase from Micrococcus lysodeikticus (E.C. 3.6.1.3.), a complex oligomeric protein, induces extensive conformational changes in it. In this report, we describe some physicochemical properties of the enzyme forms obtained. They can be summarized as follows. (1) The subunit stoichiometry of the enzyme is altered by the absorption and desorption process since most of the forms obtained are defective in gamma and delta subunits. An important reduction in the molar proportion of alpha subunit is also observed; (2) the fluorescence spectra of the different forms show progressive tyrosine residues which roughly correspond to the extent and strength of the interaction existing before elution of the enzyme; (3) circular dichroism measurements reveal changes of the secondary structure of the F1-ATPase undergoing an increase in alpha-helical content; (4) the ordered, active forms eluted from the hydrophobic chromatography columns are less stable than the native protein, as shown by dialysis experiments. These results while supporting the use of hydrophobic chromatography as a simplified model of membrane-membrane protein interaction, also indicate the need for caution in its application to the purification of complex membrane proteins.     

Glycoproteins in bacterial membranes. In vivo labeling of the sugar portion of energy-transducing ATPase and a low-molecular-weight fraction fromMicrococcus lysodeikticus membranes

The energy-transducing ATPase and a low-molecular-weight fraction ofMicrococcus lysodeikticus membranes incorporated¹⁴C label fromd-[U-¹⁴C]glucose fed to the bacteria in synthetic medium. The specific radioactivity of the sugar portion of the ATPase and low-molecular-weight fraction was, respectively, 2.65 and 2.88 times that of their amino acids. Glucose and mannose in approximately equimolar amounts were identified as the main sugars of the glycoprotein ATPase, thus confirming previous structural studies. Glucose, galactose, and mannose (1:1:2) were identified as the main sugars of the low-molecular-weight glycopeptides. These results confirm and extend the notion that glycoprotein are constituents of prokaryotic membranes.     

Biosynthetic approach to the structure of F1-ATPase (BF1 factor) from Micrococcus lysodeikticus membranes: in vivo labeling of the polypeptides and study of their relationship

The F₁-ATPase or BF₁ factor was purified from Micrococcus lysodeikticus substrain B grown in a synthetic medium in the presence of tritiated amino acids. When analyzed in sodium dodecyl sulfate-7% polyacrylamide gels, the fresh purified preparation contained α, β, γ subunits (referred as the intrinsic subunits) and two other polypeptides (designated as X and component of relative mobility 1.0) whose status as subunits remains to be established. This overall polypeptide composition was similar to that of the F₁-ATPase isolated from the same strain grown in complex medium (J. Carreira, J. M. Andreu, M. Nieto, and E. Muñoz., 1976 Mol. Cell. Biochem.10, 67–76). The distribution of ³H-labeled amino acids into purified F₁-ATPase and its constituent polypeptides under different stages of growth was used to investigate the biosynthetic relationship between the different polypeptides. The incorporation of amino acids into purified BF₁ factor was slower than that of cytoplasmic and other membrane proteins. In isotope-dilution and chase experiments, F₁-ATPase showed one of the slowest rates of decay of the incorporated label. These results point out that F₁-ATPase of M. lysodeikticus undergoes slower turnover than the overall cytoplasmic and membrane proteins. Pulse and chase experiments allowed us to conclude that the α, β, γ subunits and the components of relative mobility 1.0 are independent with differences in their turnover and therefore do not bear any apparent relation as precursors-products. The two major subunits represent seemingly the “core” of ATPase, the β subunit behaving like the most stable component. On the other hand, the γ subunit appears to be synthesized independently from this α + β complex.     

Immunochemistry of membrane F1-Adenosine triphosphatase fromMicrococcus lysodeikticus. Role of the alpha subunit

The membrane-bound ATPase activity from two substrains ofMicrococcus lysodeikticus, designated as A and B, was inhibited by antibodies raised against the two forms of purified F₁-ATPase. Form B of the enzyme, which behaved as a poorer immunogen than form A, also showed less reactivity as an antigen, independent of the physical state of the F₁-ATPase form. Antibodies were raised against the two major subunits ( and ) isolated fromM. lysodeikticus F₁-ATPase form A, which was the most stable form of the enzyme. Anti-(-subunit) serum strongly inhibited the ATPase activity of membrane-bound ATPase but showed little inhibition of the purified, soluble F₁-ATPase. The anti-(-subunit) serum inhibited the soluble F₁-ATPase, but to a lesser extent than the membrane-bound enzyme. In any event, the effect of anti- antibodies on the membrane-bound ATPase was smaller than that of anti- antibodies. It was postulated that the subunit ofM. lysodeikticus F₁-ATPase plays an essential and regulatory role in the expression of the immunochemical properties of the protein.     

Influence of the alpha-, beta- and gamma-subunits of the energy-transducing adenosine triphosphates from Micrococcus lysodeikticus in the immunochemical properties of the protein and in their reconstitution studied by a radioimmunoassay method.

We developed a sensitive and specific radioimmunoassay of the energy-transducing adenosine triphosphatase (F1-ATPase, EC 3.6.1.3) of Micrococcus lysodeikticus and extended the assay to the alpha-, beta- and gamma-subunits of the enzyme. We isolated these subunits and studied cross-reactions. We found the immunochemical properties of alpha- and beta-subunits to differ, and gamma-subunits showed an intermediate behaviour between that of alpha- and beta-subunits. Our findings indicate that each subunit of M. lysodeikticus F1-ATPase has its own identity and that conformational antigenic determinants and/or co-operative antigenic sites-arise from subunit assembly. Equimolecular amounts of alpha- and beta-subunits (up to three copies of each) reconstituted partially the immunochemical properties of the ATPase molecule, and addition of 2 mol of gamma-subunit per mol of alpha 3 beta 3 complex improved reconstitution. Our findings describe the first reconstitution of biological activity of this ATPase by assembly of the isolated subunits, and provide support for earlier proposals on the stoicheiometry of the alpha 3 beta 3 gamma 2 type for M. lysodeikticus F1-ATPase. The radioimmunoassay method affords opportunities to study the homologies between different energy-transducing ATPases and their constituent polypeptides before the primary structure of these complex proteins has been determined.     

羧基化多壁碳纳米管、混合盐及其复合胁迫对水稻幼苗生理特性的影响

将水稻（Oryza sativa L.）幼苗悬浮培养于含有羧基化多壁碳纳米管MWCNTs-COOH（0、2.5、5.0、10.0 mg/L）、50 mmol/L混合盐（1NaCl：9Na₂SO₄：9NaHCO₃：1Na₂CO₃）,以及MWCNTs-COOH+混合盐的复合溶液中,10 d后检测叶片生理生化指标变化,研究MWCNTs-COOH复合盐碱胁迫对水稻幼苗的毒性及生态风险。结果显示,与对照组相比,MWCNTs-COOH单一组诱导下水稻叶片O₂^·-和H₂O₂的产生不明显,而混合盐组和混合盐+MWCNTs-COOH复合组均诱导了O₂^·-和H₂O₂产物的大量累积。MWCNTs-COOH与混合盐复合后,加剧了O₂^·-和H₂O₂的累积,并有明显的浓度效应。活性氧（ROS）作为信号分子在一定程度上诱导了各处理组部分抗氧化酶（SOD、CAT、POD、APX）活性的升高;与混合盐组相比,低浓度混合盐+MWCNTs-COOH复合组中叶绿素a和胡萝卜素含量呈一定程度的升高;MWCNTs-COOH与混合盐复合后,抑制了叶片中可溶性糖（SS）和脯氨酸（Pro）的合成,致使相对电导率（REC）和丙二醛（MDA）含量显著升高。上述抗氧化酶活性及叶绿素a和胡萝卜素含量的升高对缓解水稻叶片氧化损伤、维持正常的光合电子传递及对过剩光能的热耗散是有益的,是水稻幼苗重要的防御机制。本研究表明MWCNTs-COOH单一处理在一定程度上诱导了水稻叶片的氧化胁迫和应激响应,与混合盐复合后加剧了叶片的氧化胁迫和应激损伤。