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N Nelson

Publications and source records attributed to N Nelson.

At least 163 records · Page 9Linked to original sources

Structure and targeted mutagenesis of the gene encoding 8-kDa subunit of photosystem I from the cyanobacterium Synechocystis sp. PCC 6803.

Photosystem I reaction center of the cyanobacterium Synechocystis sp. PCC 6803 contains seven different polypeptide subunits. The subunit with a molecular mass of about 8 kDa was isolated, and the sequence of its amino-terminal residues was determined. Oligonucleotide probes corresponding to this sequence were used to isolate the gene encoding this subunit. The gene, termed as psaE, codes for a polypeptide with a mass of 8075 Da. It is present as a single copy in the genome and is transcribed as a monocistronic messenger. The amino acid sequence of the 8-kDa subunit deduced from the gene sequence shows high homology with the deduced amino acid sequence of subunit IV of photosystem I from spinach. The DNA fragment sequenced in these studies also contains two other unidentified major open reading frames. A stable deletion mutation for the psaE gene was generated by transforming Synechocystis sp. PCC 6803 with a cloned DNA in which the psaE gene for 8-kDa subunit was replaced by a gene conferring resistance to kanamycin. The mutant strain shows minor differences in growth under photoautotrophic conditions and in the photosystem I activity in comparison to the wild type.

Amino Acid Sequence↗

Insertional inactivation of the gene encoding subunit II of photosystem I from the cyanobacterium Synechocystis sp. PCC 6803.

The cyanobacterium Synechocystis sp. PCC 6803 carries out oxygenic photosynthesis analogous to higher plants. Its photosystem I contains seven different polypeptide subunits. The cartridge mutagenesis technique was used to inactivate the psaD gene which encodes subunit II of photosystem I. A mutant strain lacking subunit II was generated by transforming wild type cells with cloned DNA in which psaD gene was interrupted by a gene conferring kanamycin resistance. The photoautotrophic growth of mutant strain is much slower than that of wild type cells. The membranes prepared from mutant cells lack subunit II of photosystem I. Studies on the purified photosystem I reaction center revealed that the complex lacking subunit II is assembled and is functional in P700 photooxidation but at much reduced rate. Therefore, subunit II of photosystem I is required for efficient function of photosystem I.

Chlorophyll↗

H+-translocating ATPase in Golgi apparatus. Characterization as vacuolar H+-ATPase and its subunit structures.

Golgi apparatus was prepared from rat liver, and enzymatic properties and the subunit structure of the H+-ATPase were characterized. GTP (and also ITP) was found to drive H+-transport with about 20% of the initial velocity as that of ATP. Bafilomycin, a specific inhibitor for vacuolar H+-ATPase, inhibited the activity at 2.5 nM. The H+-ATPase was completely inhibited in the cold in the presence of MgATP (5 mM) and NaNO3 (0.1 M). The cold inactivation of the H+-ATPase resulted in release of a set of polypeptides from Golgi membrane, with molecular masses almost identical to that of the hydrophilic sector of chromaffin granule H+-ATPase (72, 57, 41, 34, and 33 kDa). Three of these polypeptides (72, 57, and 34 kDa), cross-reacted with antibodies against the corresponding subunits of the chromaffin granule H+-ATPase. A counterpart of the 39-kDa hydrophobic component of chromaffin granule H+-ATPase was identified in the membrane, but no 115-kDa component was found. Hence, the Golgi H+-ATPase shows typical features of vacuolar H+-ATPase, in relatively low substrate specificity, its response to inhibitors, inactivation by cold treatment in the presence of MgATP, and subunit composition judged by antibody cross-reactivity.

Adenosine Diphosphate↗

Expression of the four subunits of the Torpedo californica nicotinic acetylcholine receptor in Saccharomyces cerevisiae.

Yeast expression vectors were constructed containing complementary DNA encoding the alpha-, beta-, gamma-, and delta-subunits of the Torpedo californica nicotinic acetylcholine receptor under the control of the Saccharomyces cerevisiae alcohol dehydrogenase promoter. All four plasmids were integrated into the yeast genome of a single yeast cell. The resulting yeast strain synthesized polypeptides novel to yeast that had the molecular weights and antigenic properties similar to the authentic T. californica receptor alpha-, gamma, and delta-subunits. The beta-subunit polypeptide could not be detected in this yeast strain, even though the poly(A)+ RNA from this strain contained all the information necessary for the expression of functional acetylcholine receptors in Xenopus laevis oocytes. The replacement of the beta-subunit mRNA 5'-untranslated leader and its N-terminal signal sequence by the corresponding alpha-subunit sequences, however, resulted in the expression of the beta-subunit polypeptide in yeast grown at 5 degrees C.

Animals↗

An improved formula for the judgement of intrathecally produced IgG in the presence of blood brain barrier damage.

In order to develop a better formula for discrimination between transsudated and intrathecally produced IgG, we studied the relation between cerebrospinal fluid (CSF) IgG, serum IgG, CSF albumin, and serum albumin for patients with blood brain barrier (BBB) damage. From this study we derived a new formula, IgG extended index = (CSF IgG/serum IgG)/(CSF albumin/serum albumin). Applying this index on 152 patients with BBB damage, but not expected intrathecal IgG synthesis, yielded 14 patients (9.2%) with increased (greater than 1.24) IgG extended index. In comparison, 'conventional' IgG index was increased in 34 of the patients (22%). The diagnostic sensitivity for 68 patients with multiple sclerosis (MS), did not differ significantly between the two formulas (81 and 87%, respectively). A formula for the estimation of 24 hours IgG production was also developed, but this formula was not better than IgG index. Children (0.5-16 yr) had similar IgG patterns as adults.

Adolescent↗

Lysosomal H+-translocating ATPase has a similar subunit structure to chromaffin granule H+-ATPase complex.

Subunit structure of the lysosomal H+-ATPase was investigated using cold inactivation, immunological cross-reactivity with antibodies against individual subunits of the H+-ATPase from chromaffin granules and chemical modification with N,N'-dicyclohexyl[14C]carbodiimide. The lysosomal H+-ATPase was irreversibly inhibited when incubated at 0 degrees C in the presence of chloride or nitrate and MgATP. Inactivation in the cold resulted in the release of several polypeptides (72, 57, 41, 34 and 33 kDa) from the membrane, which had the same electrophoretic mobility as the corresponding subunits of chromaffin granule H+-ATPase. Cross-reactivity of antibodies revealed that the 72, 57 and 34 kDa polypeptides were immunologically identical to the corresponding subunits of chromaffin granule H+-ATPase. Dicyclohexylcarbodiimide, which inhibits proton translocation in the vacuolar ATPase, predominantly labeled two polypeptides of 18 and 15 kDa, which compose the membrane sector of the enzyme. These results suggest that the lysosomal H+-ATPase is a multimeric enzyme, whose subunit structure is similar to the chromaffin granule H+-ATPase. The subunit structure of other vacuolar H+-ATPases, revealed by cold inactivation and immunological cross-reactivity, is also presented.

Adrenal Glands↗

The progenitor of ATP synthases was closely related to the current vacuolar H+-ATPase.

The gene encoding the proteolipid of the vacuolar H+-ATPase of yeast was cloned and sequenced. The deduced amino acid sequence of the yeast protein is highly homologous to that of the proteolipid from bovine chromaffin granules. In contrast to other membrane proteins the transmembrane segments of the bovine and yeast proteolipids were much more conserved than the hydrophilic parts. The fourth transmembrane segment, which contains the DCCD-binding site, was conserved 100%. Comparison of vacuolar and eubacterial proteolipids revealed a homology which pointed to a common ancestral gene that underwent gene duplication to form the vacuolar proteolipids. Additional support for this notion came from the amino acid sequences of subunits involved in the catalytic sectors of archaebacterial ATP synthase and plant and yeast vacuolar H+-ATPases, which reveal extensive sequence homology. Slight, but significant, homology between the archaebacterial and eubacterial ATP synthases was observed. These observations might suggest that the progenitor of ATP synthases was closely related to the present vacuolar H+-ATPases.

Amino Acid Sequence↗

Cold inactivation of vacuolar proton-ATPases.

Incubation of the reconstituted H+-ATPase from chromaffin granules on ice resulted in inactivation of the proton-pumping and ATPase activities of the enzyme. Inactivation was dependent on the presence of Mg2+, Cl-, and ATP during the incubation at low temperature. Approximately 1 mM ATP, 1 mM Mg2+, and 200 mM Cl- were required for maximum inactivation. Incubation for about 10 min on ice was required to achieve 50% inactivation. A much smaller decline in activity was observed when the enzyme was incubated at room temperature with the same chemicals. Inactivation in the cold resulted in the release of five polypeptides from the membrane with apparent molecular masses of 72, 57, 41, 34, and 33 kDa on sodium dodecyl sulfate gels. Three of the polypeptides of 72, 57, and 34 kDa were identified as subunits of vacuolar H+-ATPases by antibody cross-reactivity. Similar results were obtained with several other vacuolar H+-ATPases including those from plant sources. It was concluded that the catalytic sector of the enzyme is released from the H+-ATPase complex by cold treatment, resulting in inactivation of the enzyme.

Adenosine Triphosphate↗

Correspondence of minor subunits of plant mitochondrial F1ATPase to F1F0ATPase subunits of other organisms.

In addition to two major alpha- and beta-subunits, the soluble oligomycin-insensitive F1ATPase purified from sweet potato root mitochondria contains four different minor subunits of gamma (Mr = 35,500), delta (Mr = 27,000), delta' (Mr = 23,000), and epsilon (Mr = 12,000) (Iwasaki, Y., and Asashi, T. (1983) Arch. Biochem. Biophys. 227, 164-173). Among these minor subunits, the delta-subunit specifically cross-reacted with an antibody against the delta-subunit of maize mitochondrial F1 which contains only three minor gamma-, delta- and epsilon-subunits like F1ATPases from other organisms, indicating that the delta'-subunit is an extra subunit of sweet potato F1 which is absent in the maize F1. All of the four minor subunits of sweet potato F1 were purified and their N-terminal amino acid sequences of 30-36 residues were determined. The N-terminal sequence of gamma-subunit was homologous to those of the gamma-subunits of bacterial F1 and mammalian mitochondrial F1. The N-terminal sequence of the delta-subunit was homologous to those of the delta-subunits of bacterial F1, chloroplast CF1, and oligomycin sensitivity conferring protein of bovine mitochondrial F1F0. A sequence homology was also observed between the sweet potato epsilon-subunit and the epsilon-subunit of bovine mitochondrial F1. The N-terminal sequence of the delta'-subunit did not show any significant sequence homology to known protein sequences. These subunit correspondences place plant mitochondrial F1 at an unique position in the evolution of F1ATPase.

Amino Acid Sequence↗

A conserved gene encoding the 57-kDa subunit of the yeast vacuolar H+-ATPase.

The peripheral (catalytic) sector of vacuolar H+-ATPases contains five different polypeptides denoted as subunits A-E in order of decreasing molecular masses from 72 to 33 kDa. The gene encoding subunit B (57 kDa) of yeast vacuolar H+-ATPase was cloned on a 5-kilobase pair genomic DNA fragment and sequenced. Four open reading frames were identified in the sequenced DNA. One of them encodes a protein of 504 amino acids with a calculated Mr of 56,557. Hydropathy plot revealed no apparent transmembrane segments. Southern analysis demonstrated that a single gene encodes this polypeptide in the yeast genome. The amino acid sequence exhibits extensive identity with the homologous protein from the plant Arabidopsis (77%). This polypeptide also contains regions of homology with the alpha subunits of H+-ATPases from mitochondria, chloroplasts, and bacteria. However, less similarity was detected when it was compared with the beta subunits of those enzymes. The implication of these phenomena on the evolution of proton pumps is discussed.

Amino Acid Sequence↗

Structure, molecular genetics, and evolution of vacuolar H+-ATPases.

Proton-ATPases can be divided into three classes denoted as P-, F-, and V-ATPases. The P-ATPases are evolutionarily distinct from the F- and V-type ATPases which have been shown to be related, probably evolved from a common ancestral enzyme. Like F-ATPases, V-ATPases are composed of two distinct structures: a catalytic sector that is hydrophilic in nature and a hydrophobic membrane sector which functions in proton conduction. Recent studies on the molecular biology of vacuolar H+-ATPases revealed surprising findings about the evolution of pronon pumps as well as important clues for the evolution of eukaryotic cells.

Amino Acid Sequence↗

Cloning and nucleotide sequence analysis of genes coding for the major chlorophyll-binding protein of the moss Physcomitrella patens and the halotolerant alga Dunaliella salina.

Two clones have been isolated from a genomic library of the moss Physcomitrella patens and a cDNA library of the halotolerant green alga Dunaliella salina. The isolates contain genes coding for the major light-harvesting chlorophyll-a/b-binding protein (CAB) in the photosystem II (PSII) light-harvesting complex (LHCII). The 2544-bp insert of the moss genomic clone contains the complete CAB-coding region and 5' and 3' flanking sequences. The coding region contains an intron of 359 bp which is spanned by a pair of 9-bp perfect direct repeats. There are two CCAAT boxes and five enhancer-like elements related to (G)TGGTTTAAA(G) (Weiher et al., 1983) residing in the intron. Comparisons of the moss cab gene with sequences of light-inducible genes of higher plants reveal homologous and repeated sequences similar to the enhancer element in the 5' region upstream from the TATA and CCAAT boxes thought to be responsive to light inducibility. The 1256-bp algal cDNA contains the complete CAB-coding sequence, a 170-bp 5'-nontranslated region, and a 264-bp 3'-nontranslated region. While the overall homology in the nontranslated regions is low between the cab gene of the moss and that of the alga, the 3'-nontranslated regions of the two contain some sequences that are conserved among the cab genes in higher plants. The deduced amino acid sequences of these two clones are highly conserved except for the N-terminal region. Their hydropathic plots are very similar and both possess three hydrophobic segments that are likely alpha-helical transmembrane segments. The proposed CAB transit peptide sequence of the alga is divergent from that of the moss or higher plants, suggesting that they may have evolved from different origins. Southern blot analysis shows that the cab genes in the moss and the alga, as in higher plants, are encoded by a number of homologous genes constituting a multigene family.

Amino Acid Sequence↗

Distribution of Thylakoid Proteins between Stromal and Granal Lamellae in Spirodela : Dual Location of Photosystem II Components.

We have quantified the lateral distribution of 12 thylakoid proteins of Spirodela oligorrhiza by immunoblot analysis of detergent-derived granal and stromal lamellae. The immunological, ultrastructural, cytochemical, and biophysical measurements each indicated the expected overall separation of photosystem II (PSII) and photosystem I (PSI) components; however, certain proteins were not completely localized to one lamellar fraction. The apoproteins of the light harvesting chlorophyll a/b complex, subunit 1 of PSI and the components of the PSII reaction center (the 32 kilodalton, D2, and cytochrome b(559) proteins) were dually located between granal and stromal lamellae. Proteins associated exclusively with one of the membrane types were: in granal lamellae, the 43 and 51 kilodalton PSII proteins, and in stromal lamellae, the alpha and beta subunits of the proton ATPase.

Journal Article↗

The Q-oTc and Q-Tc interval and ionized calcium in newborns.

Ionized calcium (CaF), Q-oTc and Q-Tc intervals were determined in 31 full-term healthy newborns, 12 infants subjected to blood exchange transfusion (ET) and 10 infants with verified clinical hypocalcaemia. There was no significant correlation between CaF values and QT intervals. The large fluctuations in CaF during ET did not correspond to any similar change in Q-oTc or Q-Tc intervals. The Q-oTc interval was slightly longer in the hypocalcaemic and the blood exchanged groups compared to controls. The Q-Tc interval was somewhat longer in the ET group. The differences in mean values for QT intervals were fairly small and the ranges wide. ECG findings lack practical implications in diagnosing and following neonatal hypocalcaemia and we therefore recommend the measurement of CaF as a guideline.

Calcium↗

Plasma ionized calcium, phosphate and magnesium in preterm and small for gestational age infants.

Blood concentrations of ionized calcium, phosphate and magnesium were determined on days 1, 3 and 5 in 32 preterm infants, appropriate for gestational age, and 25 small for gestational age infants. The results were compared with those of a reference group of 31 fullterm newborns. Preterm infants had lower mean ionized calcium levels than the reference population. Ionized calcium levels were positively correlated with gestational age and postnatal age. The sickest infants had the lowest calcium levels. Intrauterine growth retardation did not seem to influence the calcium levels. Small for gestational age infants had lower phosphate values than the reference group and the values correlated with the degree of growth retardation the first day. Magnesium concentrations increased with time and did not differ significantly between the study groups and the reference group. Magnesium values did not correlate with calcium or phosphate values.

Calcium↗

Maimonides Medical Center uses workflow management system.

This optically based system integrates data, image and sound into a unified workflow management system. Running on micros, the system involves a LAN, saves space and time, and has led to a significant reduction in accounts receivable.

Accounts Payable and Receivable↗

The certification process. History and significance for addictions nursing practice.

The addictions nursing certification process will become a reality for the nurse. Ideally, this process will provide a method of ensuring quality care while maintaining self-regulation by the profession. The literature supports certification based on standards of care as a measure of that quality of care and as a voluntary mechanism selected by the practicing nurse. Many professional issues lie ahead for specialty nursing practice in the 1990s. The addictions nursing certification established by the NNSA offers a quality basis for certification implementation. The addictions nursing practice certification credential can provide the acknowledgement that the nurse has mastered a body of knowledge and skills reflective of a specialty nursing area.

Certification↗