Multiple forms of gamma-butyrobetaine hydroxylase (EC 1.14.11.1).

gamma-Butyrobetaine hydroxylase [4-trimethylaminobutyrate, 2-oxoglutarate:oxygen oxidoreductase (3-hydroxylating), EC 1.14.11.1] from human kidney was resolved into three forms by chromatofocusing. After further chromatography on an anion-exchanger, each form appeared as a single band on electrophoresis in polyacrylamide gel containing sodium dodecyl sulphate. The isoelectric points of isoenzymes 1, 2 and 3 were 5.6, 5.7 and 5.8 respectively, as estimated by isoelectric focusing. Their specific activities were 17-29 mu kat/g of protein. The concentrations of the three isoenzymes were about equal, possibly slightly lower for isoenzyme 1. The requirement for Fe2+ and the Km values for gamma-butyrobetaine and 2-oxoglutarate were about the same for the different enzyme forms. L- and D-Carnitine caused decarboxylation of 2-oxoglutarate to the same extent (8 and 29%) with the three forms. The enzyme forms had the same mass, 64 kDa, as determined by gel filtration in nondenaturing media. The same subunit mass, 42 kDa, was obtained for the multiple forms by electrophoresis in polyacrylamide gels containing sodium dodecyl sulphate. Isoenzyme 2 was resolved into two protein bands by isoelectric focusing in polyacrylamide gels containing urea. Isoenzyme 1 contained only one of these bands and isoenzyme 3 the other. The three enzyme forms of gamma-butyrobetaine hydroxylase thus appear to be dimeric combinations of two subunits differing in charge but not in size. gamma-Butyrobetaine hydroxylase from crude extracts of human, rat and calf liver was also separated into multiple forms by a chromatofocusing technique. The isoenzyme pattern was the same in human liver and kidney. The technique used to resolve the mammalian enzymes gave no evidence for the presence of multiple forms of the bacterial enzyme from Pseudomonas sp. AK 1.     

ACETYLCHOLINESTERASE OF RAT SYMPATHETIC GANGLION: MOLECULAR FORMS, LOCALIZATION AND EFFECTS OF DENERVATION

Abstract— In sucrose gradient centrifugation, acetylcholinesterase (AChE, EC 3.1.1.7.) from the rat superior cervical ganglion (SCG) has been found to contain four molecular forms, characterized by their sedimentation coefficients (4 S, 6.5 S, 10 S and 16 S). Homogenization of the ganglia in various media showed that the 4 S enzyme was readily solubilized in water whereas solubilization of the 6.5 S and 10 S forms was quantitative only in media containing Triton X-100. In order to solubilize the 16 S form, high concentrations of salt (NaCl 1 M) and detergent had to be present. AChE analysed by non-denaturing polyacrylamide gel electrophoresis separated into five bands. Although both distribution patterns were stable, i.e. each form or band preserved its characteristic sedimentation or electrophoretic migration when reanalysed, there was no 1:1 correlation between the forms isolated by sedimentation and the bands obtained by electrophoresis: one band might contain more than one form of enzyme, and conversely one form gave rise to several bands. It was therefore impossible to derive molecular weights from electrophoretic migration in non-denaturing gels. However, it could be shown that the results obtained by both methods of analysis were consistent. Acetylcholinesterase from other nervous structures was analysed: in pre- and postganglionic nerves, the main forms were 10 S and 6.5 S, with a small proportion of 4 S; the 16 S form was not detected. In other sympathetic ganglia, the distribution of forms was identical to that of the superior cervical ganglion. In rachidian ganglia, no 16 S form could be found. Following the section of the preganglionic nerve, the acetylcholinesterase activity of the superior cervical ganglion decreased by 50% in 3 days, and then rose again to about 80% of its original value after 2 weeks. These effects mainly reflected variations in the major 4 S and 10 S forms. The 16 S form, in contrast to its disappearance from denervated muscles, increased transiently during the first 2 weeks after denervation, reaching about twice its original activity. A concomitant cytochemical study of normal and denervated ganglia showed that after preganglionic denervation, AChE localized in the sympathetic neurones decreased markedly and remained low even during the recovery phase. During this period a cholinesterasic activity appeared in the perineuronal glia. Controls established that the enzyme synthetized in the glia is AChE.     

Reconstitution of the erythrocyte anion channel

Band 3, the membrane protein which mediates erythrocyte anion exchange, was purified on a concanavalin A column. Triglycerides, diglycerides, cholesteryl esters, cholesterol, and phosphatidylcholine were found to copurify. The column product gave at least two and probably three bands by isoelectric focusing. Antibodies prepared against the purified Band 3 appeared to react only with the cytoplasmic face of Band 3. Vesicles prepared with Band 3 had an accelerated uptake of SO4(2-) which could be inhibited by 2-(j'-aminophenyl)-6-methyl benzene thiazo-3', 7-disulfonic acid, a potent inhibitor of anion transport in the intact system. The possible source of this difference is discussed. Band 3 was spin labeled, probably at one specific site. The spectra showed that the spin label was highly immobilized, but no dipole-dipole interactions between spin labels on adjacent Band 3 subunits were apparent.     

Role of enzymes and identification of stage-specific proteins in developing somatic embryos of chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.)

Accumulation of proline, activities of peroxidase (POX), catalase (CAT), phenylalanine ammonia lyase (PAL) and malate dehydrogenase (MDH) were studied during different developmental stages of somatic embryos in chickpea. Callus cultures that did not form somatic embryos served as control. While increased levels of proline and POX activity were noticed in globular stages of embryos, CAT activity increased during early and late heart-shaped embryo formation indicating tissue-specific activation of these enzymes. The activity of PAL reached a peak during torpedo and cotyledonary stages of embryo development. On the other hand, MDH activity enhanced during the germination of somatic embryos inferring more requirement of energy during this stage. Electrophoretic (sodium dodecyl sulfate polyacrylamide gel electrophoresis) pattern of proteins revealed that ten bands are associated with non-embryogenic tissues, whereas 11 bands with globular, heart, torpedo and cotyledonary stages of embryo development and nine bands during the germination stage of embryos. Two extra stage-specific protein bands with molecular masses of 16 and 18 kDa appeared during globular, heart, torpedo, and cotyledonary stages. But, these bands disappeared during germination of embryos and are absent in non-embryogenic cultures. This study thus may help in the identification of proteins and the role of above enzymes during different developmental stages of somatic embryo induction and their maturation in a recalcitrant leguminous crop plant chickpea.     

不同胁迫预处理提高水稻幼苗抗寒性期间膜保护系统的变化比较

通过比较盐、冷和热激三种不同胁迫预处理提高水稻(Oryza sativa L.)抗寒性过程中膜保护系统的变化,探讨植物交叉适应机理。结果表明,水稻幼苗经不同胁迫预处理均可提高幼苗的抗寒性。与未预处理苗相比,不同胁迫(冷、热、盐)预处理之间在处理后、低温伤害后及恢复2d后的3个不同时期膜酶促和非酶促保护系统及同工酶酶谱变化各有异同,既有部分共性,也有其独特性。     

Genetic and physiological variation among bean lines resistant and susceptible to bean anthracnose

Summary Electrophoresis was used to determine genetic and or biochemical variation, if any, among bean lines resistant and susceptible to anthracnose. This was based on two enzyme systems: peroxisase and esterase. It was revealed that resistant and suceptible plants differed in their band patterns and intensities. Band intensity differences occurred mainly among monomorphic bands with higher intensities expressed by susceptible plants, while band pattern differences were expressed both by resistant and susceptible plants. These differences appeared only at certain stages of development. These stages were identified as 3 and 40 days after emergence and were considered as critical stages for screening purposes. The peroxidase isozyme A₅ and the esterase isozyme C₁ at 3 days, and the peroxidase band C₁ and esterase bands A₁ and A₂ at 40 days were important because these differences could be used as genetic/biochemical markers for screening the population for resistance. Thus, electrophoretic differences could be used as a screening aid and this could save time and effort in breeding programmes. Comparisons between inoculated and non-inoculated leaves of resistant and susceptible lines indicated that infection induced changes in both the amount and kind of peroxidases even before symptoms of the disease appeared. However, there were no specific differences between resistant and susceptible lines, indicating that resistant and susceptible lines responded to infection in the same manner.     

红豆草体细胞胚胎发生早期过氧化物酶和酯酶同工酶酶谱变化

红豆草下胚轴切段接种于含1mg/l BA,1mg/l KT的LS培养基上,通过筛选和繁殖由一块外植体而来的淡黄色愈伤组织,而得到生理状态比较一致具有较高胚性发生能力的非胚性愈伤组织,将其转移到含1mg/l BA的LS培养基上后可诱导体细胞胚眙发生。在体细胞胚胎发生早期发现过氧化物酶同工酶和酯酶同工酶酶谱均有规律性变化。过氧化物酶同工酶酶谱在胚性培养的第10天,两条明显的A_1、A_2带消失。酯酶同工酶各酶带之间酶活性比例在胚性培养过程中变化很大,培养后期酶带变得不明显或酶带数下降。说明胚性发生过程遗传信息的表达有选择性并为激素所调控。     

Appearance and distribution of dendritic cells and macrophages in dental pulp during early postnatal morphogenesis of mouse mandibular first molars

Dendritic cells and macrophages were examined in dental pulp during the postnatal development of mouse mandibular first molars, by immuno- and enzyme histochemistry. F4/80 antibody against dendritic cells and macrophages demonstrated labeled cells predominantly in and around the odontoblastic layer during tooth development from postnatal day 0 (PN0) to PN5. Labeling with Mac-1, Mac-2, and MOMA-2 antibodies against macrophages showed varied distribution patterns. Mac-1-positive cells were not detected in the dental pulp. Mac-2-positive cells appeared in the dental pulp at PN0, but not in or around the odontoblastic layer, and disappeared by PN3. A few MOMA-2-positive cells were detected in the dental pulp during the period examined. The F4/80-positive cells in and around the odontoblastic layer did not exhibit acid phosphatase or non-specific esterase activities. In addition, the F4/80-positive cells showed continued expression of Fcγ receptor, but not class II major histocompatibility complex (MHC). Other antibodies against dendritic cells (NLDC-145, MIDC-8, and 33D1) did not label the F4/80-positive cells. We concluded that the F4/80-positive and class II MHC-negative cells in and around the odontoblastic layer may be immature dendritic cells in the early stages before eruption, weaning, and crucial exposure to antigenic stimuli. They may not only act primarily as immunosurveillance cells, but also play a role in a regulatory function and differentiation of odontoblasts. Accepted: 8 June 1999     

IN VITRO MORPHOGENESIS OF AMPHIBIAN ERYTHROBLASTS

Non-mammalian vertebrate erythrocytes are flattened nucleated ellipsoids containing marginal bands (MBs) of microtubules that assemble during cellular morphogenesis. Earlier work suggested that pointed erythroid cells containing pointed MBs were intermediate stages in terminal differentiation, rather than aberrant forms, but direct evidence was lacking. Here we report on morphogenesis in individual post-cytokinetic amphibian erythroblasts in culture. Daughter cells remained adjacent in pairs, and developed pointed morphology over 1–2 h in the following sequence: (a) ends opposite the cytokinetic furrow became pointed, producing a spheroidal singly-pointed stage; (b) furrow ends usually became pointed, yielding doubly-pointed cells; (c) furrow-end points disappeared, producing a second singly-pointed stage that was flattening. Over a longer term, the single points sometimes disappeared, yielding a flattened discoid. These observations support the hypothesis that pointed cells are normal intermediates in a biogenetic program in which post-mitotic centrosomes organize MBs while occupying the singly-pointed ends of differentiating erythroblasts.     

Developmental pattern of rat intestinal brush-border enzymic proteins along the villus–crypt axis

At various postnatal stages, intestinal epithelial cells were isolated sequentially from villus tip to crypt base by successive EDTA treatments. According to the localization of marker enzymic activities, isolated cells were pooled into three cell compartments: villus (V), lower villus and upper crypt (VC) and crypt (C). Purified brush-border-membrane proteins were separated by 7.5%-polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. Enzymic activities could be assigned to some protein bands: maltase/glucoamylase (protein band 3), sucrase-isomaltase (protein bands 3 and 6), lactase (protein band 5) and alkaline phosphatase (region of protein bands 8 and 9). The findings suggest the following. (1) Sucrase-isomaltase activities appeared in compartment C at 17 days with a simultaneous increase of the pre-existing protein band 3 and appearance of a well-defined protein band in position 6; the enzymic complex remained still present in the crypt cells until adulthood. From the day 21 onwards, sucrase-isomaltase was detected in compartments VC and V. (2) Lactase was only present in the three cell compartments until day 21; at this developmental stage its activity completely disappeared from compartment C, in spite of the persistence of a weak protein band. (3) Alkaline phosphatase activity could be detected as a single peak corresponding to protein band 9 in all three cell compartments until day 21; thereafter it was replaced by two peaks of activity showing a less precise correlation with the well-defined protein bands 8 and 9. In the crypt cells of the adult rat, however, the preweaning situation, which was regularly observed, is an unexpected phenomenon. (4) Maltase and glucoamylase did not display any marked qualitative or quantitative modifications either along the villus-crypt axis or during the period of postnatal development studied. Evidence is given from the present data that each brush-border enzyme investigated has a specific developmental pattern.     

Purification and Separation of A and B Forms of N-Acetyl-β-D-Hexosaminidase from Human Aorta

Human aorta has been shown to possess multiple forms of N-Acetyl-6-D-hexosaminidase (β-2-acetamido-2-deoxy-D-glucoside-acetamido-deoxyglucohydro-lase, EC 3.2, 1.30). The enzyme was separable, by gel electrophoresis, into 2 enzymatically active bands representing A and B forms. By gel electro-focussing, A and B forms were further subdivided into at least 5 and 8 bands, respectively. The B form consisted of 4 bands (B₁) and 4 bands (B₂), which were not inactivated at 50° for 3 hr. (at pH 4.4) in the presence of serum; whereas, the 5 bands found in A form were completely inactivated. All forms of the enzyme were active towards naphthol-AS-BI-N-acetyl-β-D-glucosaminide and the corresponding galactosaminide (about one-eighth of the hydrolysis rate of the former), suggesting each single enzyme acts on both substrates. The N-acetyl-hexosaminidases of bull epididymis, by comparison, were also found to be active towards both substrates and to possess 13 bands having pis more alkaline than those of the B form of the human enzyme, By heat inactivation we found that the aortic enzyme consisted of 51% of A and 49% of B (B₁ + B₂ .). Neuraminidase had no effect on either form of the aortic preparation. Both forms were partially purified and separated by conventional methods. They required BSA for their maximal activity; the A form being more dependent BSA than the B form, With PNP-N-acetyl-β-D-glucosaminide and the corresponding galactosaminide, Km of 1.04 mH and 0.54 mM, respectively, for A form and of 1.74 and 1.48 mM, respectively, for B form were obtained. While the purified B form was stable and did not transform into other species, the purified A form gradually transformed into B form as well as into other new forms during storage at -20°.     

Expression and distribution of aggrecanases in human larynx: ADAMTS-5/aggrecanase-2 is the main aggrecanase in laryngeal carcinoma

Members of the ADAMTS family of proteases degrade proteoglycans and thereby have the potential to alter tissue architecture and regulate cellular functions. Aggrecanases are the main enzymes responsible for aggrecan degradation, due to their specific cleavage pattern. In this study, the expression status, the macromolecular organization and localization of ADAMTS-1, ADAMTS-4/aggrecanase-1 and ADAMTS-5/aggrecanase-2 in human normal larynx and laryngeal squamous cell carcinoma (LSCC) were investigated. On mRNA level, the results showed that ADAMTS-4 was the highest expressed enzyme in normal larynx, whereas ADAMTS-5 was the main aggrecanase in LSCC presenting a stage-related increase up to stage III (8-fold higher expression compared to normal), and thereafter decreased in stage IV. Accordingly, immunohistochemical analysis showed that ADAMTS-5, but not ADAMTS-4, was highly expressed by carcinoma cells. Sequential extraction revealed an altered distribution and organization of multiple molecular forms (latent, activated and fragmented forms) of the enzymes within the cancerous and their corresponding macroscopically normal laryngeal tissues, compared to the normal ones. Importantly, these analyses indicated that critical macromolecular changes occurred from the earliest LSCC stages not only in malignant parts of the tissue but also in areas that were not in proximity to carcinoma cells and appeared otherwise normal.     

Changes in levels of Proteases and Their Inhibitors during Development of the Seeds of Job's Tears (Coix lacryma-jobi L. var. Ma-yuen Stapf) and Their Localization and Interactions

Changes in levels and localization of proteases and a trypsininhibitor (JBTI) in the developing seeds of Job's tears (Coixlacryma-jobi L. var. Ma-yuen Stapf) were followed, and theirinteractions were examined. The JBTI was induced from the middlestage of development (12 DAF, i.e., 12 days after flowering)and increased until the late stage of development (24 DAF),and was localized in the germ. Two groups of proteases (A, thosewith molecular weights of 55–70 kDa; and B, those withmolecular weights greater than 94 kDa) were detected by activestaining of substrate-containing SDS-polyacrylamide gels. Thegroup of larger proteases seems to consist of three members(B₁, B₂₎ B₃, with increasing molecular weights). Band A wasobserved only in the early stage of development (until DAF 9),band B₂ persisted during all stages, while bands B, and B₃ werepresent only during early and late stages, respectively. Thegroup A proteases and one of the group B proteases (probablyB₁) were inhibited by diisopropyl-fluorophosphate (DFP), bytrypsin inhibitors from soybean and rice, and by JBTI. The proteasesthat were present in the seeds of Job's tears at a late stageseemed to be localized in the germ. (Received September 9, 1988; Accepted April 19, 1989)     

Stages of forelimb regeneration in Ambystoma maculatum.

A series of normal stages describing the regeneration of larval A. maculatum limbs after amputation through the upper arm or wrist is described. Nine discrete stages were recognized, based on external morphological and associated histological features. These stages are Initial Dedifferentiation (ID), Early Bud (EB), Medium Bud (MB), Late Bud (LB), Early Redifferentiation (ER), Notch (N), 2-Fingerbud (2-FB), 3-Fingerbud (3-FB) and 4-Fingerbud (4-FB). Similarities and differences between this and other staging systems for urodele limb regeneration are discussed. The absence of osteoclasts was a striking feature during dedifferentiation of the wrist, in contrast to their presence in large numbers during dedifferentiation of the upper arm.     

Conformational conversion of PDE5 by incubation with sildenafil or metal ion is accompanied by stimulation of allosteric cGMP binding

Phosphodiesterase-5 (PDE5) is a dimer containing a cGMP-specific catalytic domain and an allosteric cGMP-binding subdomain (GAF A) on each subunit. PDE5 exhibits three conformational forms that can be separated by Native PAGE and are denoted as Bands 1, 2, and 3 in decreasing order of mobility. A preparation comprised mainly of Band 2 PDE5 was partially converted to Band 3 PDE5 by 1 h incubation with cGMP or the PDE5-specific inhibitors sildenafil, vardenafil, or tadalafil, but not with cAMP, milrinone (PDE3-specific), or rolipram (PDE4-specific). Band 2 PDE5 was converted almost entirely to Band 3 PDE5 by overnight incubation with sildenafil at 30 °C. This time-dependent conversion was accompanied by a 7-fold increase in allosteric cGMP-binding activity, suggesting that Band 3 PDE5 is a much more active form than Band 2 PDE5 for allosteric cGMP binding. Conversion of Band 2 PDE5 to Band 3 PDE5 occurred faster by pre-incubation with cGMP, which binds to both the allosteric and catalytic sites of PDE5, than with catalytic site-specific sildenafil. Overnight incubation of a Band 2/Band 3 PDE5 mixture with EDTA caused time-dependent conversion to Band 1 PDE5 (apoenzyme), and this conversion was accompanied by a 50% loss in cGMP-binding activity. After incubation with EDTA, addition of Mn⁺⁺ or Mg⁺⁺ caused reversion of Band 1 to a Band 2/Band 3 PDE5 mixture in which Band 3 PDE5 predominated. This reversion was accompanied by a 3-fold increase in allosteric cGMP-binding activity. The combination of results implied that physiological conversion of Band 2 to Band 3 PDE5 by cGMP and/or divalent metal ion occupancy of the catalytic domain would increase allosteric cGMP binding to the enzyme. This conversion would produce a greater negative feedback effect on cGMP action by increasing sequestration of cGMP at the allosteric cGMP-binding site of PDE5 and by increasing cGMP degradation at the catalytic site of the enzyme. This conversion would also increase PDE5 inhibitor binding to the enzyme.     

Persistence and expression of circular DNAs encoding Drosophila amylase, bacterial chloramphenicol acetyltransferase, and others in Xenopus laevis embryos

We injected circular forms of several different DNAs into fertilized eggs of Xenopus laevis, and studied the persistence and expression of the injected DNAs during early embryonic development. When we injected plasmids which contained Drosophila amylase genes, the copy number of the injected DNA increased only slightly during cleavage, started to decrease at the blastula stage, then became very small at the tadpole stage. In such embryos, Drosophila amylase activity was detected at and after the gastrula stage. When we injected other kinds of circular DNAs (pX1r101A, cDm2055, pII25.1, pBR322, and pSP-64-L14), their copy number did not increase throughout the early stages. When circular plasmids that contained bacterial chloramphenicol acetyltransferase (CAT) genes were injected, their copy number usually did not increase, but sometimes, for unknown reasons, it increased extensively throughout the blastula to gastrula stages. In both cases, CAT enzyme activity started to be expressed during the blastula to gastrula stages and disappeared at the 2 day-old tadpole stage. The level of CAT enzyme activity was roughly proportional to the amount of CAT mRNA formed, and also to the copy number of injected genes. From these results, we concluded that in Xenopus embryos, exogenously-injected circular DNAs are preserved for the most part as circular DNAs, and that the increase in their copy number within the embryos is not prerequisite for the expression of their genetic information.     

Multiple cysteine proteinase forms during the life cycle of Dictyostelium discoideum revealed by electrophoretic analysis.

Proteinases of the cellular slime mould Dictyostelium discoideum have been analysed using electrophoresis on polyacrylamide gels containing gelatin (gelatin/PAGE). Multiple proteinase forms were apparent in vegetative myxamoebae, but the presence of individual enzyme forms depended on the manner in which the cells were grown. Axenic cells had a characteristic A-pattern of proteinases consisting of six bands, the most active enzymes having apparent Mr values of 51,000 and 45,000 (these have been named ddCP51 and ddCP45, respectively). Some of the proteinases were also present in the medium, the major extracellular form was ddCP42, a 42,000-Mr enzyme. Cells grown in association with bacteria had a distinct B-pattern with three main enzymes that had apparent Mr values of 48,000, 43,000 and 38,000. All of the A- and B-pattern proteinases were most active at acid pH in the presence of dithiothreitol and were inhibited by various agents such as trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E64), leupeptin and chymostatin, which inactivate cysteine proteinases. One of the enzymes, ddCP30, was identified as cysteine proteinase B which had been purified and characterized previously [North, M.J. & Whyte, A. (1984) J. Gen. Microbiol. 130, 123-134]. During starvation of axenic cells in shaken suspensions some of the vegetative proteinases disappeared, ddCP42 was released from the cells and one new enzyme with an apparent Mr of 48,000 appeared. Addition of cyclic AMP had little effect on these changes. When the axenically grown myxamoebae underwent development on filters, similar changes in band pattern were observed and the aggregation stage was characterized by the presence of three cysteine proteinase bands (apparent Mr values of 48,000, 45,000 and 43,000). Proteinases, especially ddCP42, were released from the cells and could be collected from the buffer-saturated pads which supported the filters. The results demonstrate that cysteine proteinases are present throughout growth and development of D. discoideum and that the forms present are subject to nutritional and developmental regulation.     

MULTIPLE FORMS OF CHOLINE ACETYLTRANSFERASE AND THE HIGH AFFINITY UPTAKE OF CHOLINE IN BRAIN OF DEVELOPING AND ADULT RATS

The multiple molecular forms of choline acetyltransferase (ChAT) were analysed during the postnatal development of rat brain. Changes in the sodium-dependent, high affinity uptake of [³H]choline (HAUC) and in the efficiency of conversion of labelled choline into ACh in vitro were also examined. Both mature and 7-day old brain contained three molecular forms of ChAT, with isoelectric points of pH 7.3, 7.9 and 8.3, but the immature brain appeared to contain smaller concentrations of the most basic form of the enzyme (pI = 8.3). Of the total choline uptake measured in slices of frontal cortex, adult samples exhibited a greater proportion of HAUC than 7-day samples and appeared to acetylate more efficiently the [³H]choline accumulated by high affinity uptake. This evidence suggests a basic molecular form of ChAT, appearing in rat brain during postnatal development, might be responsible for the efficient coupling of the high affinity uptake and subsequent acetylation of choline in cholinergic nerve terminals.     

Comparative electrophoretic characteristics of alkaline phosphatase isoforms in maternal blood and shaggy chorion extracts at various periods of pregnancy]

A comparative study of enzyme alkaline phosphatase (AP) of plasma and chorion placental extracts was made in various terms of pregnancy by PAAG electrophoresis. It is shown that the first three month chorion extract contained 1 thermolabile (TL) and 1 or 3 thermostable (TS) fractions of AP. In the second trimester of intrauterine development the activity of TL fraction disappeared, the number of TS forms increased. The early plasma period (6-12 weeks) was characterized by 2 TL non-placental forms, from the second half they added to 1 or 3 TS placental fractions, and before delivery TS enzyme appeared in the liver AP which had no analogue in the placenta. It is concluded that only the fastest placental TS AP forms (1 or 3) appear in maternal blood from the second trimester of gestation.     

Sequential changes in amylase isozymes during grain maturation in barley

Summary Changes in amylase isozyme patterns on polyacrylamide gels were followed during maturation in grains of Deba Abed barley. Early stage seeds contained a single, high-mobility enzyme (Band A) which had an estimated molecular weight of 4.2×10⁴ and a high activity with -limit dextrin as a substrate. It was shown, by dissection, that Band A was confined to the aleurone layer and probably represented the initial product of amylase synthesis.This form was succeeded, in mid-course, by a less mobile form (Band B), a -amylase with a molecular weight of approximately 1.3×10⁵. Late-dough stage grains contained a complex of low-mobility -amylase bands which were shown, by papain digestion, to be protein-bound forms of Band B.The changes are discussed on the basis of a unified series consisting of elaborated forms of the initial Band A type of activity.