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。     

松墨天牛取食和人为损伤对马尾松针叶部分化学物质含量的影响

为探索松墨天牛Monochamus alternatus Hope与其寄主马尾松相互作用的关系,测定了松墨天牛取食和人为损伤1天、3天、6天、10天和15天后马尾松针叶营养物质和次生代谢物质含量的变化。结果表明： 松墨天牛取食和人为损伤均能引起马尾松针叶可溶性糖和粗多糖含量的下降,但虫害处理使新叶和老叶中可溶性糖降低更多。两种方式处理后10天马尾松新叶和老叶中蛋白质含量降到最低,然后逐渐回升; 相比较而言,虫害处理蛋白质含量降幅更大。松墨天牛取食和人为损伤均引起马尾松针叶单宁和黄酮类物质含量的增加,虫害处理的针叶中单宁和黄酮类物质的含量更高。结果提示马尾松对松墨天牛的取食为害具有特殊的应激反应机制。     

马尾松受害诱导的化学物质滞后变化

通过接种马尾松毛虫Dendrolimus punctatus和人工剪叶,分析比较了不同受害方式对马尾松针叶内化学物质的影响。结果表明,害虫取食松针诱导的次生代谢物质(单宁、酚类物质)比人工剪叶处理略有增加,而营养物质-可溶性糖变化不大。对马尾松进行害虫取食为害、人工剪叶受害、未受害三种处理后,连续3年跟踪测定了松针叶内化学物质含量的变化。结果发现,无论是害虫取食为害,还是人工剪叶受害,针叶内次生代谢物质含量都减少,可溶性糖含量降低,直至一年后才恢复到原有的水平;蛋白质变化的趋势则始终是受害处理的含量比未受害处理的含量高,表明马尾松受害后诱导的化学物质变化具有滞后特性。     

BACE蛋白的表达、纯化和活性测定

在大肠杆菌中表达、纯化并重新折叠以获得有活性的酸性蛋白水解酶 (BACE蛋白 )———一种与阿尔茨海默病 (AD)发病相关的蛋白水解酶。克隆BACE活性区的表达序列到原核表达载体 pET11a中 ,经E .coliBL2 1(DE3)表达 ,从包涵体中获取蛋白质 ,电泳鉴定后经梯度反向快速折叠法重新折叠 ,柱层析分离纯化 ,得到了表达的重组可溶性BACE蛋白 ;用高效液相色谱、质谱等方法检测其对人工合成多肽底物的水解作用 ;测定了BACE蛋白的酶促动力学常数。结果表明 ,得到的重组BACE蛋白具有水解人工合成小肽底物的活性。     

蛋白质色谱的高效化技术

过去30年间,以基因工程和细胞融合技术为代表的现代生物技术的发展让治疗和诊断用蛋白质的规模化生产成为现实.与此同时,蛋白质色谱、膜分离等生物下游核心技术的创新也适应了这种发展.以蛋白质色谱为例,琼脂糖、纤维素和亲水性高分子聚合物等材料被广泛地应用于色谱介质的合成,其孔道结构得到一定的改良;离子交换色谱等技术日趋完善,新型色谱技术(如金属螯合亲和色谱等)不断涌现;色谱的生产效率通过过程集成和外场(如电场、磁场、超重力场等)的引入得以进一步提高.同时,蛋白质色谱也为基因组学、蛋白质组学等组学研究提供了有力的工具.     

外源激素对马尾松花芽分化及内源物质的影响

为探讨马尾松球花形成与植物激素水平的关系,该研究对贵州省都匀无性系种子园11年生马尾松进行不同浓度的 IAA、IBA、GA3、BAP等植物激素处理,采用考马斯亮蓝G-250染色法、蒽酮法分别对不同浓度不同激素处理后的枝条上针叶中的可溶性蛋白质和可溶性糖含量变化进行测定,并在第二年开花时对试验枝条的开花情况进行了调查。结果表明：在8－11月份,进行500 mg·L-1的BAP 处理有利于马尾松雌球花和雄球花的形成,100 mg·L-1的GA3处理有利于马尾松雌球花的形成;而GA3250 mg·L-1和GA3500 mg·L-1处理有利于马尾松雄球花的形成,IAA 250 mg·L-1对马尾松雌雄球花同枝的数量有提高作用。在10－11月份,对马尾松进行500 mg·L-1的BAP、IAA、GA3处理后,马尾松针叶内蛋白质含量变化有显著影响;在10月份时,进行BAP 100 mg·L-1处理后,其可溶性糖含量及可溶性蛋白含量均可达到极显著水平。而在8月份与10月份时,分别进行IBA 100 mg·L-1与IBA 250 mg·L-1处理后,其可溶性蛋白含量与其对照差异处于极显著水平;在11月份时,进行GA3100 mg·L-1处理后,其可溶性蛋白含量与其对照差异处于极显著水平;而在11月份时,进行IBA 500 mg·L-1处理后,其可溶性蛋白含量与对照差异处于显著水平。     

马尾松幼苗生长及生理特性对铝胁迫的响应

以马尾松幼苗为试验材料,采用水培法研究铝胁迫（Al³⁺浓度为0、0.2、0.4、0.8、1.6 mmol·L^-1）对马尾松幼苗生长及其针叶中叶绿素、渗透调节物质（可溶性糖、可溶性蛋白、脯氨酸）、丙二醛（MDA）和超氧化物歧化酶（SOD）、过氧化物酶（POD）等保护酶活性的影响,为揭示马尾松铝毒害生理机制及提高马尾松的耐铝能力提供理论依据。结果显示：当Al³⁺处理浓度为0.2 mmol·L^-1时对马尾松株高和基径生长的影响较小,但对马尾松根系生长有一定的促进作用;Al³⁺处理浓度大于0.2 mmol·L^-1时对马尾松株高、基径和根长的生长均会产生一定的抑制作用,且这种抑制作用随着Al³⁺浓度的增大而增强。马尾松针叶中叶绿素含量和SOD、POD活性均随着Al³⁺处理浓度的增加呈先上升后下降的趋势;Al³⁺处理浓度大于0.2 mmol·L^-1时马尾松针叶中可溶性糖、可溶性蛋白、脯氨酸等渗透调节物质均呈上升趋势,且随着Al³⁺浓度的升高而增大;Al³⁺处理浓度大于0.2 mmol·L^-1时马尾松针叶中MDA含量也呈上升趋势,且随着Al³⁺浓度的增大而升高,说明大于0.2 mmol·L^-1的Al³⁺处理可导致马尾松膜脂产生氧化。研究表明,马尾松幼苗具有一定的耐铝能力,在铝胁迫生境下可通过提高自身SOD和POD等保护酶的合成和主动积累脯氨酸、蛋白质和可溶性糖等渗透物质,产生适应性生理响应以维持自身的生理平衡来降低铝毒害作用。     

核糖核酸酶A超家族：不仅仅是一组降解RNA的酶

核糖核酸酶A超家族(ribonuclease A superfamily; RNase A superfamily),也称脊椎动物分泌型核糖核酸酶超家族(vertebrate secreted ribonucleases superfamily),是二十世纪蛋白质结构、酶学和分子进化领域研究最多最广泛的核糖核酸酶家族。自上世纪初期从牛胰腺中分离鉴定第一个成员以来，已从哺乳动物、两栖动物、爬行动物、鸟和鱼等几百种动物中鉴定了几千个成员。早期对该家族成员的研究不仅促进了蛋白质化学技术的发展，而且为现代生物学研究奠定了基础。目前已知人的核糖核酸酶A超家族成员包括8个典型成员(RNase 1～RNase 8)和5个非典型成员(RNase 9～RNase 13)。功能方面，曾一度以为该家族成员只具有降解核糖核酸的能力。随着血管生成素(angiogenin; RNase 5)、嗜酸性粒细胞衍生神经毒素(eosinophils-derived neurotoxin, EDN; RNase 2)、嗜酸性粒细胞阳离子蛋白(eosinophils cationic protein, ECP; RNase ...     

一株高效溶磷伯克霍尔德菌的筛选鉴定及对马尾松幼苗的促生作用

采用标准稀释平板法从马尾松根际土中分离溶磷细菌,利用钼锑抗比色法测定溶磷细菌的溶磷特性;通过分析溶磷菌的溶磷能力与发酵液pH的关系,以及液相色谱-质谱 (HPLC-MS)联用对发酵液中有机酸的测定,探究其溶磷机制;通过对接种溶磷菌马尾松盆栽苗生长、生理、土壤养分和土壤酶活性的测定,明确溶磷菌对马尾松生长和生理的影响。结果表明: 由马尾松根际土壤中共筛选到溶磷细菌16株,其中菌株WJ27溶磷效果最优,液体培养5 d时的溶磷量达411.98 mg·L^-1。经过表型观察、生理生化鉴定和系统发育树分析,发现菌株WJ27属于伯克霍尔德菌属;其对不同磷源的溶磷特性存在差异,溶磷能力依次为: Ca₃(PO₄)₂(220.85 mg·L^-1)＞AlPO₄(182.33 mg·L^-1)＞FePO₄·2H₂O(129.19 mg·L^-1)＞CaHPO₄·2H₂O (115.23 mg·L^-1)。胞外有机酸测定结果表明,该菌株通过分泌柠檬酸、丙二酸等有机酸降低发酵液中pH,进而发挥溶磷作用;盆栽试验结果表明,接种菌株WJ27对马尾松幼苗生长、生理、土壤养分和土壤酶活性有积极作用。与对照相比,接种WJ27的马尾松的苗高、主根长、侧根数量、地上部(茎、枝、叶)鲜重、干重和根系鲜重、干重分别增加了14.3%、36.9%、56.1%、44.7%、60.0%、158.3%和100.0%;叶绿素b、总叶绿素、地上部可溶性蛋白和可溶性糖、根系活力和根系可溶性蛋白分别增加了145.8%、45.2%、206.3%、59.4%、80.5%和260.0%;根系超氧化物歧化酶、过氧化物酶和地上部过氧化氢酶的活性分别提高了71.2%、197.5%和36.6%;根际土壤中速效氮、速效钾、速效磷含量和土壤脲酶、过氧化氢酶、磷酸酶活性分别较对照土壤显著增加18.1%、17.0%、11.9%和34.3%、45.5%、62.6%。说明接种WJ27可以改善土壤养分和土壤酶活性,进而促进马尾松幼苗的生长。     

改进的薄层组织电泳技术对发育过程中鸡胚腰段脊髓蛋白质变化的分析

本文介绍一种改进的结合组织化学和生物化学的电泳方法,用此方法对神经组织中微量蛋白质进行定性、定量和定位的分析。结果表明在不同发育阶段中,鸡胚腰段脊髓中绝大多数蛋白区带表现为色度上的深浅变化,少数区带随发育而丢失或新生,脊髓背腹区域的蛋白区带非常相似,而酸性糖蛋白却表现为腹部多于背部。此外,还分析了鸡胚和鸡(刍鸟)的7种组织中蛋白质区带的差异。     

Dephosphorylation of neuromodulin by calcineurin

Neuromodulin (p57, GAP-43, F1, B-50) is a major neural-specific, calmodulin binding protein found in brain, spinal cord, and retina that is associated with membranes. Phosphorylation of neuromodulin by protein kinase C causes a significant reduction in its affinity for calmodulin (Alexander, K. A., Cimler, B. M., Meirer, K. E., and Storm, D. R. (1987) J. Biol. Chem. 262, 6108-6113). It has been proposed that neuromodulin may function to bind and concentrate calmodulin at specific sites within neurons and that activation of protein kinase C causes the release of free calmodulin at high concentrations near its target proteins. It was the goal of this study to determine whether bovine brain contains a phosphoprotein phosphatase that will utilize phosphoneuromodulin as a substrate. Phosphatase activity for phosphoneuromodulin was partially purified from a bovine brain extract using DEAE-Sephacel and Sephacryl S-200 gel filtration chromatography. The neuromodulin phosphatase activity was resolved into two peaks by Affi-Gel Blue chromatography. One of these phosphatases, which represented approximately 60% of the total neuromodulin phosphatase activity, was tentatively identified as calcineurin by its requirement for Ca2+ and calmodulin (CaM) and inhibition of its activity by chlorpromazine. Therefore, bovine brain calcineurin was purified to homogeneity and examined for its phosphatase activity against bovine phosphoneuromodulin. Calcineurin rapidly dephosphorylated phosphoneuromodulin in the presence of micromolar Ca2+ and 3 microM CaM. The apparent Km and Vmax for the dephosphorylation of neuromodulin, measured in the presence of micromolar Ca2+ and 2 microM CaM, were 2.5 microM and 70 nmol Pi/mg/min, respectively, compared to a Km and Vmax of 4 microM and 55 nmol Pi/mg/min, respectively, for myosin light chain under the same conditions. Dephosphorylation of neuromodulin by calcineurin was stimulated 50-fold by calmodulin in the presence of micromolar free Ca2+. Half-maximal stimulation was observed at a calmodulin concentration of 0.5 microM. We propose that phosphoneuromodulin may be a physiologically important substrate for calcineurin and that calcineurin and protein kinase C may regulate the levels of free calmodulin available in neurons.     

The Amino Acid Sequence of the Tryptophan-Containing Subunit (α-Subunit) of Bovine Brain S-100 Protein

Abstract: The tryptophan-containing subunit (α-subunit) of bovine brain S-100 protein was purified from a S -aminoethyl derivative of S-100a protein, and its amino acid sequence was determined. The α-subunit contained 93 residues, including one tryptophan, and had a molecular weight of 10,400. The sequence shows an extensive homology (58% identity) to the sequence of another "tryptophan-free" subunit (β-subunit) found in both S-100a and S-100b protein, and has a calcium binding site characteristic of the "E-F hand" proteins, such as calmodulin or troponin C. The tryptophan residue is located at position 90 which is presumably adjacent to the C-terminal end of the α-helix following the calcium binding loop, and thus appears likely to serve as a specific probe in structure-function studies of S-100a protein.     

Molecular cloning and the complete nucleotide sequence of cDNA to mRNA for S-100 protein of rat brain.

The complete nucleotide sequence of mRNA for beta-subunit of rat brain S-100 protein was determined from recombinant cDNA clones. The sequence was composed of 1488 bp which included the 276 bp of the complete coding region, the 120 bp of the 5'-noncoding region and the 1092 bp of the 3'-noncoding region containing two polyadenylation signals. In addition, the poly(A) tail was also found. The amino acid sequence deduced from the nucleotide sequence was homologous to the amino acid sequence of bovine S-100 beta subunit except 4 residues showing species differences. From the viewpoint of evolutionary implications, the homology between the nucleotide sequence of S-100 and those of rat intestinal Ca-binding protein (ICaBP) and calmodulin (CaM) was examined. A dot-blot hybridization of poly(A) RNA from the developing rat brains using a labeled cDNA showed a rapid increase in S-100 mRNA at 10-20 postnatal days. The presence of S-100 mRNA in C-6 glioma cells is also described.     

Isolation and characterization of a calcium-binding protein derived from mRNA termed p9Ka, pEL-98, 18A2, or 42A by the newly synthesized vasorelaxant W-66 affinity chromatography.

W-66 (N-(2-aminoethyl)-N-[2-(4-chlorocinnamylamino) ethyl]-5-isoquinolinesulfonamide), a newly synthesized isoquinolinesulfonamide, was shown to have a potent vasodilatory action and calmodulin (CaM)-antagonizing action. Using the W-66 affinity chromatographic technique, we purified two Ca(2+)-binding proteins from the EGTA-soluble fraction of bovine aorta. One was CaM and the other was an acidic protein with a molecular mass of 11 kDa. It was tentatively named "calvasculin." Calvasculin was a dimeric protein. Equilibrium dialysis showed that 1 mol of calvasculin (dimer) bound to 1.98 +/- 0.30 mol of Ca2+ in the presence of 10(-3) M Ca2+. Calvasculin failed to activate Ca2+/CaM-dependent enzymes such as myosin light chain kinase, Ca2+/CaM-dependent phosphodiesterase, or Ca2+/CaM-dependent protein kinase II and to inhibit the CaM stimulation of these enzymes. The partial amino acid sequence of calvasculin revealed a high homology to the predicted protein derived from mRNA, named pEL-98, 18A2, 42A, or p9Ka. We also examined the physicochemical and biochemical properties of calvasculin. Using the antibody specific for calvasculin, we obtained evidence that calvasculin is present in abundance in bovine aorta but not in brain, lung, heart, or testis.     

Membrane lipids of human peripheral nerve and spinal cord.

Major membrane lipids were determined in specimens of human peripheral nerve (cauda equina) and spinal cord of 10 subjects aged 20-70 years. The same lipids were also assayed in myelin from the same tissues isolated with two different procedures and in myelin of cauda equina from 3 subjects aged 17-91 years isolated with a third method. The concentrations (mean and standard deviation) of phospholipids were 90 +/- 11 and 96 +/- 9 nmol/g fresh weight; of cholesterol 70 +/- 15 and 101 +/- 16; of cerebroside 19 +/- 3 and 41 +/- 7; of sulfatide 10 +/- 1 and 11 +/- l; and of gangliosides 0.80 +/- 0.08 and 0.40 +/- 0.05 N in cauda equina and spinal cord, respectively. The proportion of ethanolamine phosphoglyceride was lower and that of sphingomyelin higher in cauda equina than in spinal cord. The myelin of peripheral nerve and spinal cord contained almost the same proportions of lipids as the whole tissue. The protein-bound sialic acid content was 3-fold higher than the lipid-bound sialic acid content in cauda myelin. The fatty acid patterns of choline, ethanolamine, inositol and serine phosphoglycerides of spinal cord and its myelin, were very similar to those of cerebral white matter, while the phosphoglycerides of cauda equina had higher proportions of monoenoic acids and lower proportions of polyunsaturated fatty acids. The fatty acid patterns of sphingomyelin, cerebroside and sulfatide of spinal cord were similar to those of cerebral white matter, while those of cauda equina contained significantly more saturated fatty acids. This suggests that the lipid and fatty acid compositions of peripheral nerve are particularly suitable for the formation of a tightly packed myelin membrane which can be a powerful shield against infections and other injuries.     

Does aluminum inhibit pollen germination via extracellular calmodulin?

The effect of aluminum (Al) on pollen germination and its mechanism of action were investigated. Pollen germination and pollen tube elongation were inhibited by Al at pH 4.5. This inhibitory effect was reversed by the addition of purified calmodulin (CaM), whereas neither the calcium binding-protein S-100 nor Al chelator citric acid at the same concentrations had any obvious effect on Al-inhibited pollen germination. The presence of either the membrane-impermeable CaM inhibitor anti-CaM antiserum or Ca2+ chelator EGTA completely suppressed the effect of exogenous CaM. These results indicate the involvement of extracellular calmodulin in the short-term effects of Al on pollen germination and pollen tube elongation.     

Calmodulin X (Ca2+)4 is the active calmodulin-calcium species activating the calcium-, calmodulin-dependent protein kinase of cardiac sarcoplasmic reticulum in the regulation of the calcium pump

Calcium-, calmodulin-dependent phosphorylation of cardiac sarcoplasmic reticulum increases the rate of calcium transport. The complex dependence of calmodulin-dependent phosphoester formation on free calcium and total calmodulin concentrations can be satisfactorily explained by assuming that CaM X (Ca2+)4 is the sole calmodulin-calcium species which activates the calcium-, calmodulin-dependent, membrane-bound protein kinase. The apparent dissociation constant of the E X CaM X (Ca2+)4 complex determined from the calcium dependence of calmodulin-dependent phosphoester formation over a 100-fold range of total calmodulin concentrations (0.01-1 microM) was 0.9 nM; the respective apparent dissociation constant at 0.8 mM free calcium, 1 mM free magnesium with low calmodulin concentrations (0.1-50 nM) was 2.60 nM. These results are in good agreement with the apparent dissociation constant of 2.54 nM of high affinity calmodulin binding determined by 125I-labelled calmodulin binding to sarcoplasmic reticulum fractions at 1 mM free calcium, 1 mM free magnesium and total calmodulin concentration ranging from 0.1 to 150 nM, i.e. conditions where approximately 98% of the total calmodulin is present as CaM X (Ca2+)4. The apparent dissociation constant of the calcium-free calmodulin-enzyme complex (E X CaM) is at least 100-fold greater than the apparent dissociation constant of the E X CaM X (Ca2+)4 complex, as judged from non-saturation 125I-labelled calmodulin binding at total calmodulin concentrations of up to 150 nM, in the absence of calcium.     

Probing the role of calmodulin in Al toxicity in maize

The role of calmodulin on Al toxicity was studied in two maize (Zea mays L.) inbred lines, Cat 100-6 (Al-tolerant) and S 1587-17 (Al-sensitive). Increasing levels of Al induced the release of malate at similar rate by roots of both genotypes, while the exudation of citrate, a stronger Al-binding compound, was 3.5 times higher in Cat 100-6 seedlings exposed to 16.2x10(-6) Al(3+) activity. The calmodulin inhibitor trifluoperazine significantly reduced the root growth in both genotypes, mimicking the main effect of Al. However, when Cat 100-6 and S 1587-17 seedlings were challenged with Al in conjunction with trifluoperazine, no further reduction in root growth or any other effect of Al toxicity was observed. The rate of Al-induced citrate exudation by both genotypes was not affected by treatment with trifluoperazine or calmidazolium, another calmodulin inhibitor. The Al(3+) interaction with cytoplasmic CaM was estimated using models for the binding of Al(3+) and Mg(2+) with CaM and physiological concentrations of citrate, CaM, InsP(3), ATP, ADP, Al(3+) and Mg(2+). In this simulation, Al(3+) associated with citrate and InsP(3), but not with CaM. We conclude that calmodulin is not relevant to the physiological processes leading to the Al tolerance in maize, nor is it a primary target for Al toxicity.     

Affinity chromatography of brain cyclic nucleotide phosphodiesterase using 3-(2-pyridyldithio)propionyl-substituted calmodulin linked to thiol-sepharose

[3-(2-Pyridylthio)propionyl]calmodulin (PDP-CaM), an activated thiol derivative of calmodulin (CaM), was synthesized. Preparations of this derivative containing an average of 2.8 mol of substituent/mol of protein activated purified cyclic nucleotide phosphodiesterase in a manner indistinguishable from that of native CaM. PDP-CaM was covalently coupled to free thiol-Sepharose 4B through formation of a stable mixed disulfide bond for use in affinity chromatography. The binding capacity of the disulfide-linked CaM-Sepharose for phosphodiesterase activity was proportional to substituent level up to 4 mg of CaM/mL of gel; the total capacity of the gel for binding phosphodiesterase was 4 times that of CNBr-coupled CaM-Sepharose. Quantitative recovery was achieved by desorption of both ligand and bound proteins with a reducing agent. The thiolated CaM derivative was then separated from phosphodiesterase by rapid gel filtration; the overall recovery of phosphodiesterase activity was greater than 70%. Preparations of homogeneous enzyme in good yield were obtained after a second chromatography step on CaM-Sepharose. Binding and recovery of phosphodiesterase activity were entirely reproducible, since each preparation of affinity gel was used only once. As it permits separation of interacting species in free solution, this general method may be useful with other ligands for increasing yields from affinity chromatography, particularly when dissociation of molecules in their matrix-bound conformation may be difficult to achieve.     

Isolation of S-100 binding proteins from brain by affinity chromatography

S-100-binding proteins, and calmodulin-binding proteins were isolated from S-100- and calmodulin-depleted bovine brain extract by Ca2+-dependent affinity chromatography using S-100- and calmodulin-coupled Sepharose columns respectively. The majority of the protein (80 to 90%) including calcineurin that bound to S-100 also bound to calmodulin and vice versa, suggesting both proteins may regulate common targets. However these two regulatory proteins also bind few other proteins specific for each. These include cyclic nucleotide phosphodiesterase, 55k, and 220k proteins for calmodulin and 24k, 42k, and 90k proteins for S-100. Certain proteins also specifically bound to S-100 both in Ca2+-dependent and independent ways. In glial cells S-100 protein may replace calmodulin in regulating Ca2+-influenced functions.