侵蚀退化地植被恢复过程中芒萁对土壤可溶性有机碳的影响

林下植被是生态系统的重要组分。通过对比分析红壤侵蚀区植被恢复过程中,林下有无芒萁覆盖地的土壤可溶性有机碳(DOC,Dissolved Organic Carbon)含量及其与地下根系生物量、地上植被淋溶液DOC含量的关系。结果表明:林下植被芒萁覆盖增加了地上叶片和地下根系生物量,土壤DOC含量及储量也显著增加(P0.05),芒萁覆盖对表层土壤(0—20cm)DOC的影响大于深层土壤(20—100cm)(P0.05);相关分析结果表明,林下芒萁覆盖地土壤DOC储量与细根生物量的垂直变化呈显著的正相关关系(P0.05),且随植被恢复年限的增加相关性显著增加,地下根系的垂直分布直接影响各土层DOC储量。不同植被恢复时期,林下芒萁覆盖地土壤DOC与鲜叶(马尾松+芒萁)和枯落物(马尾松+芒萁)淋溶液DOC均呈显著的正相关关系(P0.01),而林下裸露地土壤DOC仅与鲜叶(马尾松)淋溶液DOC呈显著的相关性(P0.01),林下芒萁覆盖地相对于裸露地枯落物淋溶液对土壤DOC储量的影响大于鲜叶。植被恢复过程中芒萁覆盖地土壤微生物生物量碳和微生物熵显著高于林下裸露地。因此,在植被恢复进程中,芒萁能够提供更多底物参与土壤物质与养分循环,对土壤DOC的贡献较大,为侵蚀区马尾松林恢复提供了重要的养分再吸收来源;同时芒萁覆盖增加了微生物活性,促进了微生物对土壤DOC的同化作用,提高了微生物碳源的利用率,对土壤有机碳的积累起着重要的作用。     

红壤侵蚀区芒萁对土壤微生物群落结构的影响

土壤微生物是反映土壤质量状况的重要指标,研究侵蚀地植被恢复后土壤微生物群落结构的变化对深入认识土壤质量的演变具有重要意义。对比分析了未治理地(Y0)、治理13年(Y13)和31年(Y31)的马尾松林(Pinus massoniana)林下芒萁(Dicranopteris dichotoma)覆盖地(NRd)、去除芒萁覆盖地(Rd)与林下裸地(CK)土壤微生物生物量和群落结构差异,结果表明:林下裸地土壤微生物生物量碳(MBC)、微生物生物量氮(MBN)和总微生物磷脂脂肪酸量(总PLFAs)的含量均显著低于芒萁覆盖地,且去除芒萁4个月后,MBC和总PLFAs均有降低趋势,表明芒萁覆盖对土壤微生物生物量具有重要影响;林下芒萁覆盖地土壤革兰氏阳性菌(GP)、革兰氏阴性菌(GN)、丛植菌根真菌(VAM)、真菌(Fungi)、放线菌(ACT)的PLFAs含量显著高于林下裸地(Y13例外),去除芒萁4个月后,各值均有有接近林下裸地的趋势;芒萁覆盖地真菌/细菌的比值(F/B)均显著高于林下裸地(P0.05),芒萁覆盖地革兰氏阳性菌/革兰氏阴性菌的比值(GP/GN)、饱和直链脂肪酸/单不饱和脂肪酸的比值(sat/mono)和(cy17:0+cy19:0ω8c)/(16:1ω7c+18:1ω7c)(cy/pre)显著小于林下裸地(P0.05),去除芒萁4个月后,芒萁覆盖地土壤cy/pre显著升高(P0.05)(Y13例外),意味着芒萁覆盖地土壤生态系统更稳定,土壤的养分可利用性更高,微生物生物量和群落结构更丰富,活性更强;皮尔逊相关分析和冗余分析发现,土壤理化性质与土壤微生物生物量和群落结构关系密切,土壤C/N、p H和氮素水平是调控芒萁覆盖下土壤微生物生物量和群落结构的主要生态因子。     

川西高山森林林窗对季节性冻融期土壤氮动态的影响

为了解高山森林林窗对土壤氮动态的影响,于2012—2013年在川西高山冷杉原始林大、中、小林窗以及林下采集了4个关键时期(初冻期、深冻期、初融期和融化末期)的土样,测定其铵态氮、硝态氮、微生物生物量氮和可溶性有机氮含量。结果表明:各林窗内土壤铵态氮、硝态氮含量在融化末期显著高于其他3个关键时期;土壤微生物生物量氮在初融期最低,土壤可溶性有机氮含量在深冻期最低,而这两者含量在初冻期均最高;土壤硝态氮含量占土壤矿质氮总量的67.26%~83.59%;冬季林窗通过改变土壤微环境进而引起氮素组分的改变,林窗大小与可溶性有机氮含量呈显著正相关;土壤温度与铵态氮、硝态氮及可溶性有机氮含量呈显著正相关;冻结深度与硝态氮和可溶性有机氮呈显著负相关。经过季节性冻融期,小林窗和林下土壤具有更高的矿质氮和可溶性有机氮,为生长季内植被与土壤微生物奠定了良好的生长条件。     

松材线虫对马尾松林土壤微生物生物量及酶活性的影响

以感染松材线虫病的马尾松林土壤作为研究对象,探索不同程度松材线虫病感染对马尾松林土壤理化性质、微生物生物量和土壤酶活性的影响。结果表明:随松材线虫病危害程度的加重,总碳、总磷、总钾、可溶性有机碳、铵态氮、硝态氮、有效磷和含水量呈现升高趋势,而p H、Ca、Mg、可溶性有机氮、微生物生物量碳(MBC)和微生物生物量氮(MBN)显著降低;同时,蔗糖酶、脲酶、纤维素酶和多酚氧化酶酶活性随着感染程度的加重而趋于下降,而酸性磷酸酶和蛋白酶活性则显著升高。结果表明:土壤脲酶、蔗糖酶、纤维素酶和多酚氧化酶4种酶活性与含水量、铵态氮、硝态氮、可溶性有机碳、总磷、总钾、有效磷、总碳等理化指标呈显著负相关,而与可溶性有机氮、p H、Mg、Ca含量呈显著正相关;酸性磷酸酶和蛋白酶酶活性与含水量、铵态氮、硝态氮、可溶性有机碳、总钾呈显著正相关,而与可溶性有机氮、p H、Mg和Ca呈显著负相关;另外,MBC和MBN与酸性磷酸酶或蛋白酶呈显著负相关,而MBC和MBN与蔗糖酶、纤维素酶或多酚氧化酶呈显著正相关;因此,松材线虫的侵染改变了松林土壤的理化性质,引起土壤微生物群落结构、生物量和土壤酶活性的变化,这些指标可用于指示和评价松材线虫侵染对土壤质量的影响。     

红壤侵蚀区芒萁对土壤可溶性有机质光谱特征的影响

可溶性有机质(DOM)是森林生态系统能量循环的主要载体,在碳循环过程中发挥着重要的作用。为了深入了解植被恢复后土壤DOM的变化和结构特征,在典型红壤侵蚀区福建省龙岩市长汀县河田镇选取不同恢复年限的马尾松(Pinus massoniana)林为研究对象,利用光学技术对比分析了不同恢复年限(0年,13年,31年)马尾松林保留芒萁(Dicranopteris dichotoma)覆盖地、去除芒萁覆盖地和林下裸地土壤DOM光谱特征。结果表明:未治理地(Y0)、恢复13年(Y13)和恢复31年(Y31)马尾松林芒萁覆盖地土壤可溶性有机碳(DOC)含量分别是林下裸地的7.61倍、4.83倍和5.47倍,去除芒萁一年后,土壤DOC的含量显著下降,但仍分别是林下裸地的1.84倍、4.12倍和4.73倍;芒萁覆盖地土壤DOM的芳香化指数(AI)和腐殖化指数(HIX)均显著高于林下裸地,而波长在250 nm和365 nm处的紫外可见光光度值之比(E2:E3)的趋势与之相反,去除芒萁一年后,AI和HIX降低显著,表明芒萁覆盖地土壤DOM腐殖化和芳香化程度更高,分子量更大;林下裸地DOM红外光谱中特征峰明显不如林下芒萁覆盖地丰富,其含有更多的羟基、羧酸类,以及碳水化合物中的烷氧基等结构简单、易迁移的物质,去除芒萁一年后,DOM红外光谱特征峰无明显变化,表明芒萁是土壤DOM数量和结构的主要影响因素,而这种影响是一个长期缓慢的过程。从DOM光谱分析结果可知,芒萁覆盖下土壤DOM的分子量更大,结构更复杂,易于被土壤胶粒吸附,维持其化学稳定,利于土壤有机碳积累。由此可见,芒萁在土壤有机碳积累过程中具有积极的作用。     

恢复年限、林下植被及季节对马尾松林土壤氮转化的影响

侵蚀红壤区植被恢复能有效防治土壤侵蚀,改善生态环境。提高侵蚀退化地土壤氮矿化潜力、增加氮有效性是改善贫瘠土壤植被生长发育的关键途径,对恢复侵蚀地生态系统具有重要意义。采用顶盖埋管培养法研究了不同恢复年限(Y0、Y16、Y34))不同芒萁处理的马尾松林土壤净氮矿化量和净氮矿化速率的季节变化特征,分析了植被恢复年限、林下植被覆盖及季节变化对土壤氮矿化的交互影响。结果表明,植被恢复能使侵蚀退化地土壤养分条件得到改善。不同恢复年限马尾松林净氮矿化最高值出现在夏秋季,而在春季为负值。植被恢复能使土壤净氮矿化量显著增加,且净氮矿化过程以氨化作用为主。净氨化速率与净矿化速率具有相似的季节变化,硝化速率随着恢复年限增加季节变化减小。林下裸露地净氮矿化量及速率低于芒萁覆盖地,且去除芒萁可以降低净氮矿化量及速率。方差分析表明,恢复年限、季节变化及其交互作用能显著影响土壤净氮矿化量及矿化速率(P0.001),而芒萁处理未能达到显著水平(P0.05)。马尾松林土壤氮转化过程季节变化明显,林分管理应按季节变化进行,林下芒萁覆盖对侵蚀退化地马尾松林土壤氮恢复具有重要作用。     

天山林区不同类型群落土壤氮素对冻融过程的动态响应

季节性冻融过程对北方温带森林土壤氮素的转化与流失具有重要影响,但不同类型群落对冻融过程响应的差异尚不明确。通过在林地、草地、灌丛上设置系列监测样地,采用原位培养的方法,利用林冠遮挡形成的自然雪被厚度差异,监测分析了冻融期天山林区不同群落表层土壤(0—15 cm)的氮素动态及净氮矿化速率间的差异。结果表明:(1)不同类型群落土壤的铵态氮(NH+4-N)含量、微生物量氮(MBN)含量基本与土壤(5 cm)温度呈正相关,深冻期林地土壤铵态氮含量低于其他群落类型而硝态氮含量高于其他群落类型;(2)硝态氮(NO-3-N)为天山林区季节性冻融期间土壤矿质氮的主体,占比达78.4%。灌丛土壤硝态氮流失风险较大,融化末期较融化初期灌丛土壤硝态氮含量下降了64.6%;(3)冻融时期对整体氮素矿化速率影响显著,群落类型对氨化速率影响显著;(4)天山林区土壤氮素在冻结期主要以氮固持为主。通过揭示不同类型群落土壤氮素对冻融格局的响应,能够助益于对北方林区冬季土壤氮素循环的认识。     

黄土丘陵区植被恢复对土壤可溶性氮组分的影响

为探究黄土丘陵地区人工植被恢复对土壤氮素养分累积与有效性的影响,研究分析了植被恢复15年刺槐、柠条、刺槐侧柏混交、刺槐山桃混交以及荒草地土壤可溶性氮组分含量及其垂直分布特征。结果表明,与耕地相比,植被恢复显著提高了0—30 cm土壤可溶性氮组分含量,这也使0—30 cm土壤可溶性氮组分密度显著增加,可溶性有机氮密度增幅表现为柠条(262.2%)刺槐(232.8%)刺槐山桃混交、刺槐侧柏混交(34.5%)荒草地(-21.5%),硝态氮密度整体表现为柠条刺槐刺槐山桃混交荒草地刺槐侧柏混交,增幅为7.9%—182.8%,铵态氮密度以刺槐山桃混交增幅最大(110.3%),荒草地最小为2.6%。可溶性有机氮、硝态氮占全氮的比例以刺槐最高,分别提升了2.4倍和0.6倍,铵态氮占全氮的比例以刺槐山桃混交最高,提升了1.0倍。可溶性氮组分受微生物量碳氮的影响大于有机质和全氮,微生物量氮与可溶性氮组分的相关性优于微生物量碳,硝态氮对土壤有机质、全氮和微生物量碳氮的变化最为敏感。综上,植被恢复能够提高土壤可溶性氮组分含量、密度及其占全氮比例,增加土壤氮的有效性,以刺槐、柠条提升效果最好。     

湿地土壤微生物功能多样性及碳氮组分对长期氮输入的响应

氮输入对湿地生态系统碳氮循环具有重要影响,研究湿地土壤微生物功能多样性及碳氮组分对氮输入的响应,对于明确湿地土壤碳氮循环微生物驱动机制具有重要意义。依托长期野外氮输入模拟试验,利用Biolog-ECO微平板技术,分析不同浓度氮输入:N1(6 g N m^-2 a^-1)、N2(12 g N m^-2 a^-1)和N3(24 g N m^-2 a^-1)对湿地土壤表层(0-15 cm)和亚表层(15-30 cm)微生物碳源代谢活性、功能多样性和碳氮组分的影响。结果表明:N2处理显著提高了亚表层土壤微生物碳源代谢活性和McIntosh指数,N3处理显著降低了表层土壤微生物Shannon指数和Shannon-evenness指数。随氮输入浓度增加湿地表层土壤微生物对糖类的利用率显著降低,N3处理表层土壤微生物对胺类的利用率以及亚表层土壤微生物对醇类的利用率显著提高。N1处理显著提高了湿地表层土壤全氮和微生物量碳含量;N2、N3处理显著提高了土壤铵态氮、硝态氮含量;N3处理显著降低了土壤pH值。湿地土壤pH、总碳、溶解性有机碳含量是影响微生物碳源代谢活性和功能多样性的重要因素,土壤溶解性有机碳、铵态氮、全氮含量、含水率是影响微生物碳源利用变化的主要因子。     

氮添加对苏北沿海杨树人工林土壤活性有机碳库的影响

大气氮沉降成为目前全球性的环境问题之一,氮的沉降可能显著影响森林土壤碳循环过程。从2012年5月起,对东台林场3种林龄(5、9、15年生)黑杨派无性系I-35杨(Populus deltoides CL‘35’)人工林进行野外模拟氮沉降试验,探讨氮沉降对不同林龄杨树人工林土壤活性有机碳的影响。经过1年施氮试验后,5和9年生杨树人工林的土壤微生物生物量碳随着氮沉降水平的增加呈现出先增加后减少的趋势;而15年生林分在不同氮处理下,土壤微生物生物量碳均有所增加;3种林龄在不同氮处理下土壤可溶性有机碳含量随着氮浓度的增加而增加。土壤微生物生物量碳与可溶性有机碳之间以及这二者与土壤全氮、微生物生物量氮、可溶性有机氮、铵态氮、硝态氮之间存在显著相关。试验表明,氮沉降可能增加土壤活性有机碳含量,从而影响杨树人工林土壤碳动态。     

Expression of the Catalytic Subunit of Ouabain-Sensitive H+,K+-ATPase in Rat Skin Epidermis

A comparative localization of Na⁺,K⁺-ATPase and ouabain-sensitive H⁺,K⁺-ATPase in rat skin was performed using in situ RNA hybridization and immunohistochemistry. Na⁺,K⁺-ATPase was predominantly detected in the basal layer of the epithelium, whereas the ouabain-sensitive H⁺,K⁺-ATPase, in the granular and prickle cell layers. The genes of these ATPases are thus expressed in epithelial cells at different stages of their development. The hypothesis was advanced that the ouabain-sensitive H⁺,K⁺-ATPase is involved in maintaining the skin pH value. The probes specific to the mRNAs of the full-size -subunit of the ouabain-sensitive H⁺,K⁺-ATPase and its truncated form were used to establish a similar distribution of both mRNA variants in skin.     

1H and13C-NMR analysis of four pentasaccharides from urine of blood-group A and B,Leb adults. Confirmation of the structure of two new oligosaccharides: Fuc(α1–2)[GalNAc(α1–3)]Gal(β1–3)[Fuc(α1–4)]Glc and Fuc(α1–2)[Gal(α1–3)]Gal(β1–3)[Fuc(α1–4)]Glc

The major pentasaccharides Fuc(1-2)[GalNAc(1-3)]Gal(1-4)[Fuc(1-3)]Glc and Fuc(1-2) [Gal(1-3)]Gal(1-4)[Fuc(1-3)]Glc, which are normally present in the urine of bloodgroup A Le^b and B Le^b healthy subjects, were each found to be contaminated by a minor component when analysed by¹H-NMR. The determination of these structures, Fuc(1-2) [GalNAc(1-3)]Gal(1-3)[Fuc(1-4)]Glc and Fuc(1-2) [Gal(1-3)]Gal(1-3)[Fuc(1-4)]Glc, was based on the results of methylation analysis and¹H/¹³C-NMR spectroscopy.Abbreviations HPLC high performance liquid chromatography - GLC gas liquid chromatography - NMR nuclear magnetic resonance - COSY correlation spectroscopy - Gal d-galactopyranose - GalNAc 2-acetamido-2-deoxy-d-galactopyranose - Glc d-glucopyranose - Fuc l-fucopyranose - LNDFH I lacto-N-difucohexaose I (Le^b determinant     

Assignment of the1H- and13C-NMR spectra of eight oligosaccharides of the lacto-N-tetraose and neotetraose series

The assignment of the 13C- and 1H-NMR spectra of eight oligosaccharides of the lacto-N-tetraose and neotetraose series was obtained from homonuclear and heteronuclear correlation spectroscopy. These analyses were performed on the following compounds: 1. Gal beta 1-3GlcNAc beta 1-3Gal beta 1-4Glc; 2. NeuAc alpha 2-3Gal beta 1-3GlcNAc beta 1-3Gal beta 1-4Glc; 3. Gal beta 1-3[NeuAc alpha 2-6]GlcNAc beta 1-3Gal beta 1-4Glc; 4. NeuAc alpha 2-3Gal beta 1-3[NeuAc alpha 2-6]GlcNAc beta 1-3Gal beta 1-4Glc; 5. NeuAc alpha 2-3Gal beta 1-3[Fuc alpha 1-4]GlcNAc beta 1-3Gal beta 1-4Glc; 6. Fuc alpha 1-2Gal beta 1-3[NeuAc alpha 2-6]GlcNAc beta 1-3Gal beta 1-4Glc; 7. Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc; 8. NeuAc alpha 2-6Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc.     

Elucidation of Structure of Lipid A from the Marine Gram-Negative Bacterium Pseudoalteromonas haloplanktis ATCC 14393T

The chemical structure of lipid A, from the marine -proteobacterium Pseudoalteromonas haloplanktis 14393, a main product of lipopolysaccharide hydrolysis (1% AcOH), was determined using chemical methods and NMR spectroscopy. The lipid A was shown to be -1,6-glucosaminobiose 1,4-diphosphate acylated with two (R)-3-hydroxyalkanoic acid residues at C3 and C3 and amidated with one (R)-3-hydroxydodecanoyl and one (R)-3-dodecanoyloxydodecanoyl residue at N2 and N2, respectively.     

(Na + K)-ATPase activity of crude homogenates of rat skeletal muscle as estimated from their K-dependent 3-O-methylfluorescein phosphatase activity

A highly sensitive fluorimetric assay using 3-O-methylfluorescein phosphate as substrate was used in the determination of K⁺-dependent phosphatase activity in preparations of rat skeletal muscle. The gastrocnemius muscle was chosen because of mixed fibre composition. Crude, detergent treated homogenate was used so as to avoid loss of activity during purification. K⁺-dependent phosphatase activities in the range 0.19–0.37 μmol · (g wet weight)⁻¹ · min⁻¹ were obtained, the value decreasing with age and K⁺-deficiency. Complete inhibition of the K⁺-dependent phosphatase was obtained with 10⁻³ M ouabain. Using a KSCN-extracted muscle enzyme the intimate relation between K⁺-dependent phosphatase activity and (Na⁺ + K⁺)-activated ATP hydrolysis could be demonstrated. A molecular activity of 620 min⁻¹ was estimated from simultaneous determination of K⁺-dependent phosphatase activity and [³H]ouabain binding capacity using the partially purified enzyme preparation. The corresponding enzyme concentration in the crude homogenates was calculated and corresponded well with the number of [³H]ouabain binding sites measured in intact muscles or biopsies hereof.     

Annular and non-annular binding sites on the (Ca + Mg)-ATPase

Quenching of the fluorescence of the (Ca²⁺ + Mg²⁺)-ATPase purified from muscle sarcoplasmic reticulum can be used to measure relative binding constants of hydrophobic compounds to the phospholipid-protein interface. We show that the binding constant for cholesterol is considerably less than that for phosphatidylcholine, so that cholesterol is effectively excluded from the phospholipid annulus around the ATPase. However, dibromocholestan-3β-ol causes quenching of the fluorescence of the ATPase, and so has access to other, non-annular sites. We suggest that these non-annular sites could be at protein/protein interfaces in ATPase oligomers. Oleic acid can bind at the phospholipid/protein interface, although its binding constant is less than that for a phosphatidylcholine, and it can also bind at the postulated non-annular sites. The effects of these compounds on the activity of the ATPase depend on the structure of the phospholipid present in the systems.     

Structure of a new nonasaccharide isolated from human milk: VI2Fuc,V4Fuc,III3Fuc-p-lacto-N-hexaose

The structure of a new nonasaccharide isolated from human milk has been investigated. By using methylation analysis, FAB-MS and¹H-and¹³C-NMR spectroscopy as basic methods of structural investigation, this oligosaccharide was identified as VI²--Fuc,V⁴-Fuc,III³--Fuc-p-lacto-n-hexaose: Fuc1-2Gal1-3[Fuc1-4]GlcNAc1-3Gal1-4[Fuc1-3]GlcNAc1-3Gal1-4Glc.Abbreviations COSY correlation spectroscope - DP degree of polymerisation - FAB-MS fast atom bombardment-mass spectrometry - HPLC high performance liquid chromatography - NMR nuclear magnetic resonance - GLC gas-liquid chromatography     

Structural Peculiarities of Na+,K+-ATPase Isozymes from the Calf Brain

Functionally active preparations of Na⁺,K⁺-ATPase isozymes from calf brain that contain catalytic subunits of three types (1, 2, and 3) were obtained using two approaches: a selective removal of contaminating proteins by the Jorgensen method and a selective solubilization of the enzyme with subsequent reconstitution of their membrane structure by the Esmann method. The ouabain inhibition constants were determined for the isozymes. The real isozyme composition of the Na⁺ pump from the grey matter containing glial cells and the brain stem containing neurons was determined. The plasma membranes of glial cells were shown to contain mainly Na⁺,K⁺-ATPase of the 11 type and minor amounts of isozymes of the 22(1) and the 31(2) type. The axolemma contains 21 and 31 isozymes. A carbohydrate analysis indicated that 11 enzyme preparations from the brain grey matter substantially differ from the renal enzymes of the same composition in the glycosylation of the 1 isoform. An enhanced sensitivity of the 3 catalytic subunit of Na⁺,K⁺-ATPase from neurons to endogenous proteolysis was found. A point of specific proteolysis in the amino acid sequence PNDNR⁴⁹² Y⁴⁹³ was localized (residue numbering is that of the human 3 subunit). This sequence corresponds to one of the regions of the greatest variability in 1-, 2-, 3-, and 4-subunits, but at the same time, it is characteristic of the 3 isoforms of various species. The presence of the 3 isoform of tubulin (cytoskeletal protein) was found for the first time in the high-molecular-mass Na⁺,K⁺-ATPase 31 isozyme complex isolated from the axolemma of brain stem neurons, and its binding to the 3 catalytic subunit was shown.     

Cloning and characterization of a Na<Superscript>+</Superscript>/H<Superscript>+</Superscript> antiporter gene of the moderately halophilic bacterium <Emphasis Type="Italic">Halobacillus aidingensis</Emphasis> AD-6<Superscript>T</Superscript>

A gene encoding a Na(+)/H(+) antiporter was obtained from the genome of Halobacillus aidingensis AD-6(T), which was sequenced and designated as nhaH. The deduced amino acid sequence of the gene was 91% identical to the NhaH of H. dabanensis, and shared 54% identity with the NhaG of Bacillus subtilis. The cloned gene enable the Escherichia coli KNabc cell, which lack all of the major Na(+)/H(+) antiporters, to grow in medium containing 0.2 M NaCl or 10 mM LiCl. The nhaH gene was predicted to encode a 43.5 kDa protein (403 amino acid residues) with 11 putative transmembrane regions. Everted membrane vesicles prepared from E. coli KNabc cells carrying NhaH exhibited Na(+)/H(+) as well as Li(+)/H(+) antiporter activity, which was pH-dependent with the highest activity at pH 8.0, and no K(+)/H(+) antiporter activity was detected. The deletion of hydrophilic C-terminal amino acid residues showed that the short C-terminal tail was vital for Na(+)/H(+) antiporter activity.     

Fluorescent labeling of (Na + K)-ATPase by 5-iodoacetamidofluorescein

5-Iodoacetamidofluorescein (5-IAF) covalently labels dog kidney (Na⁺ + K⁺)-ATPase with approximately 2 moles incorporated per mole of enzyme. ATPase and K⁺-phosphatase activities are fully retained after reaction, and the kinetic parameters for Na⁺, K⁺, Mg²⁺, ATP and p-nitrophenyl phosphate are likewise not significantly affected. The fluorescence of the bound 5-IAF is increased by ATP, Na⁺, and Mg²⁺, and decreased by K⁺. These fluorescence changes likely reflect ligand-induced stabilization of the E₁ or E₂ states of the enzyme.