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Article: Cloning of promoter-active DNA sequences from Chainia (NCL 82.5.1) in Escherichia coli.

Chauthaiwale, V M / Deshpande, V V

1994 Volume 4, Issue 3-4, Page(s) 173–175

Abstract: The ability of Escherichia coli cells to recognise and use the regulatory signals of genes from Chainia (NCL 82-5-1) was determined in vivo using gene fusions. DNA fragments from Chainia were cloned in E. coli using the promoter probe plasmid pJAC4. Four ...

Abstract	The ability of Escherichia coli cells to recognise and use the regulatory signals of genes from Chainia (NCL 82-5-1) was determined in vivo using gene fusions. DNA fragments from Chainia were cloned in E. coli using the promoter probe plasmid pJAC4. Four of the randomly selected recombinants exhibited varying strengths of promoter activity as assessed by the concentration of ampicillin required to kill 50% of the colonies (LD50 values) and by beta-lactamase activity. The origin of the inserts was confirmed by colony hybridization of clones with labelled genomic DNA of Chainia. The beta-lactamase activity of recombinant colonies was substantially higher (> 10-fold) than that of colonies transformed with pJAC4 without any insert. The results show that a few Chainia DNA sequences are recognized by E. coli as transcription initiation signals for the expression of beta-lactamase gene, inspite of the high guanine/cytosine content of the Chainia genome.
MeSH term(s)	Actinomycetales/genetics ; Ampicillin/pharmacology ; Cloning, Molecular ; DNA Probes ; DNA, Bacterial/genetics ; Drug Resistance, Microbial/genetics ; Escherichia coli/genetics ; Gene Expression ; Kanamycin ; Nucleic Acid Hybridization ; Plasmids ; Promoter Regions, Genetic ; Regulatory Sequences, Nucleic Acid ; Transformation, Bacterial ; beta-Lactamases/genetics ; beta-Lactamases/metabolism
Chemical Substances	DNA Probes ; DNA, Bacterial ; Kanamycin (59-01-8) ; Ampicillin (7C782967RD) ; beta-Lactamases (EC 3.5.2.6)
Language	English
Publishing date	1994-05
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	59230-4
ISSN	1872-8081 ; 0951-6433
ISSN (online)	1872-8081
ISSN	0951-6433
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Article: Molecular cloning and expression of the xylanase gene from Chainia in Escherichia coli.

Chauthaiwale, V M / Deshpande, V V

FEMS microbiology letters

1992 Volume 78, Issue 2-3, Page(s) 265–270

Abstract: A complete genomic library of Chainia was constructed in coliphage lambda vector gt10 and was screened for the xylanase gene using an 18-mer mixed oligonucleotide probe corresponding to a six-amino acid sequence of low molecular mass Chainia xylanase. ... ...

Abstract	A complete genomic library of Chainia was constructed in coliphage lambda vector gt10 and was screened for the xylanase gene using an 18-mer mixed oligonucleotide probe corresponding to a six-amino acid sequence of low molecular mass Chainia xylanase. Inserts from 11 putative clones, showing hybridization with the oligonucleotide probe at medium stringency, were subcloned in pUC8 and screened for xylanase gene expression using anti-xylanase antibodies. The restriction map of the insert (1.4 kb) from one of the four immunopositive clones (PVX8) showing detectable xylanase activity was constructed. The xylanase activity of PVX8 was not induced by IPTG or xylan. Reorientation of the insert by directional cloning into pUC9 had no effect on the xylanase activity suggesting that an indigenous promoter from Chainia is responsible for the xylanase activity.
MeSH term(s)	Actinomycetales/genetics ; Bacteriophage lambda/genetics ; Cloning, Molecular ; Escherichia coli/genetics ; Gene Expression ; Genes, Fungal ; Genetic Vectors ; Glycoside Hydrolases/genetics ; Restriction Mapping ; Xylan Endo-1,3-beta-Xylosidase
Chemical Substances	Glycoside Hydrolases (EC 3.2.1.-) ; Xylan Endo-1,3-beta-Xylosidase (EC 3.2.1.32)
Language	English
Publishing date	1992-12-01
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	752343-9
ISSN	1574-6968 ; 0378-1097
ISSN (online)	1574-6968
ISSN	0378-1097
DOI	10.1016/0378-1097(92)90038-p
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Article: Chemical modification of xylanase from alkalothermophilic Bacillus species: evidence for essential carboxyl group.

Chauthaiwale, J / Rao, M

Biochimica et biophysica acta

1994 Volume 1204, Issue 2, Page(s) 164–168

Abstract: ... Deshpande, V, Hinge, J. and Rao, M. (1990) Biochim. Biophys. Acta 1041, 172-177). This finding and present ... The role of carboxyl group in the catalytic action of xylanase (M(r) 35,000 ... activity with a second order rate constant of 3300 M-1 min-1. The spectrophotometric analysis at 340 nm ...

Abstract	The role of carboxyl group in the catalytic action of xylanase (M(r) 35,000) from an alkalothermophilic Bacillus sp. was delineated through kinetic and chemical modification studies using Woodward's Reagent K. The kinetics of inactivation indicated that one carboxyl residue was essential for the xylanase activity with a second order rate constant of 3300 M-1 min-1. The spectrophotometric analysis at 340 nm revealed that the inhibition was correlated with modification of 24 carboxyl residues. In the presence of protecting ligand, modification of one carboxyl group was prevented. The pH profile showed apparent pK values of 5.2 and 6.4 for the free enzyme and 4.9 and 6.9 for enzyme-substrate complex. The pH dependence of inactivation was consistent with the modification of carboxyl group. The kinetic analysis of the modified enzyme showed similar Km and lower kcat values than the native enzyme indicating that catalytic hydrolysis and not the substrate binding was affected by chemical modification. The chemical modification of xylanase from alkalothermophilic Bacillus revealed the presence of tryptophans in the active site (Deshpande, V, Hinge, J. and Rao, M. (1990) Biochim. Biophys. Acta 1041, 172-177). This finding and present studies demonstrated the experimental evidence for the participation of carboxyl as well as tryptophan groups as essential residues of xylanase from alkalothermophilic Bacillus sp.
MeSH term(s)	Amino Acids/analysis ; Bacillus/enzymology ; Carboxylic Acids/chemistry ; Glycoside Hydrolases/chemistry ; Hydrogen-Ion Concentration ; Isoxazoles ; Kinetics ; Xylan Endo-1,3-beta-Xylosidase
Chemical Substances	Amino Acids ; Carboxylic Acids ; Isoxazoles ; N-ethyl-5-phenylisoxazolium-3'-sulfonate (3BN309R76H) ; Glycoside Hydrolases (EC 3.2.1.-) ; Xylan Endo-1,3-beta-Xylosidase (EC 3.2.1.32)
Language	English
Publishing date	1994-02-16
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	60-7
ISSN	1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650 ; 0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
ISSN (online)	1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650
ISSN	0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
DOI	10.1016/0167-4838(94)90004-3
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Article ; Online: The effect of a diiodothyronine mimetic on insulin sensitivity in male cardiometabolic patients: a double-blind randomized controlled trial.

van der Valk, Fleur / Hassing, Carlijne / Visser, Maartje / Thakkar, Purav / Mohanan, Anookh / Pathak, Kaushal / Dutt, Chaitanya / Chauthaiwale, Vijay / Ackermans, Mariette / Nederveen, Aart / Serlie, Mireille / Nieuwdorp, Max / Stroes, Erik

PloS one

2014 Volume 9, Issue 2, Page(s) e86890

Abstract: Background and aims: Obesity and its associated cardiometabolic co-morbidities are increasing worldwide. Since thyroid hormone mimetics are capable of uncoupling the beneficial metabolic effects of thyroid hormones from their deleterious effects on ... ...

Abstract	Background and aims: Obesity and its associated cardiometabolic co-morbidities are increasing worldwide. Since thyroid hormone mimetics are capable of uncoupling the beneficial metabolic effects of thyroid hormones from their deleterious effects on heart, bone and muscle, this class of drug is considered as adjacent therapeutics to weight-lowering strategies. This study investigated the safety and efficacy of TRC150094, a thyroid hormone mimetic. Materials and methods: This 4-week, randomized, placebo-controlled, double-blind trial was conducted in India and The Netherlands. Forty subjects were randomized at a 1:1 ratio to receive either TRC150094 dosed at 50 mg or placebo once daily for 4 weeks. Hyperinsulinemic euglycemic clamp and (1)H-Magnetic Resonance Spectroscopy (MRS) were performed before and after treatment. Results: At baseline, subjects were characterized by markedly impaired hepatic and peripheral insulin sensitivity. TRC150094 dosed 50 mg once daily was safe and well tolerated. Hepatic nor peripheral insulin sensitivity improved after TRC150094 treatment, expressed as the suppression of Endogenous Glucose Production from 59.5 to 62.1%; p = 0.477, and the rate of glucose disappearance from 28.8 to 26.4 µmol kg(-1)min(-1), p = 0.185. TRC150094 administration did not result in differences in fasting plasma free fatty acids from 0.51 to 0.51 mmol/L, p = 0.887 or in insulin-mediated suppression of lipolysis from 57 to 54%, p = 0.102. Also, intrahepatic triglyceride content was unaltered. Conclusion: Collectively, these data show that, in contrast to the potent metabolic effects in experimental models, TRC150094 at a dose of 50 mg daily does not improve the metabolic homeostasis in subjects at an increased cardiometabolic risk. Further studies are needed to evaluate whether TRC150094 has beneficial effects in patients with more severe metabolic derangement, such as overt diabetes mellitus and hypertriglyceridemia. Trial registration: clinicaltrials.gov NCT01408667.
MeSH term(s)	Adult ; Diiodothyronines/pharmacology ; Glucose Clamp Technique ; Humans ; India ; Insulin Resistance/physiology ; Magnetic Resonance Spectroscopy ; Male ; Metabolic Syndrome/metabolism ; Middle Aged ; Netherlands ; Statistics, Nonparametric ; Thyronines/pharmacology
Chemical Substances	Diiodothyronines ; Thyronines ; TRC150094 (B93608MTGW)
Language	English
Publishing date	2014-02-21
Publishing country	United States
Document type	Journal Article ; Randomized Controlled Trial ; Research Support, Non-U.S. Gov't
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0086890
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Article: Production and purification of extracellular D-xylose isomerase from an alkaliphilic, thermophilic Bacillus sp

Chauthaiwale, J / Rao, M

Applied and environmental microbiology. Dec 1994. v. 60 (12)

1994

Abstract: An alkaliphilic, thermophilic Bacillus sp. (NCIM 59) produced extracellular xylose isomerase at pH 10 and 50 degree C by using xylose or wheat bran as the carbon source. The distribution of xylose isomerase as a function of growth in comparison with ... ...

Abstract	An alkaliphilic, thermophilic Bacillus sp. (NCIM 59) produced extracellular xylose isomerase at pH 10 and 50 degree C by using xylose or wheat bran as the carbon source. The distribution of xylose isomerase as a function of growth in comparison with distributions of extra- and intracellular marker enzymes such as xylanase and beta-galactosidase revealed that xylose isomerase was truly secreted as an extracellular enzyme and was not released because of sporulation or lysis. The enzyme was purified to homogeneity by ammonium sulfate precipitation followed by gel filtration, preparative polyacrylamide gel electrophoresis, and ion-exchange chromatography. The molecular weight of xylose isomerase was estimated to be 160,000 by gel filtration and 50,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, indicating the presence of three subunits. The enzyme is most active at pH 8.0 and with incubation at 85 degrees C for 20 min. Divalent metal ions Mg2+, Co2+, and Mn2+ were required for maximum activity of the enzyme. The Km values for D-xylose and D-glucose at 80 degrees C and pH 7.5 were 6.66 and 142 mM, respectively, while Kcat, values were 2.3 X 10(2) s-1 and 0.5 X 10(2) s-1, respectively.
Keywords	thermophilic bacteria ; isomerases ; purification ; enzyme activity ; xylose ; glucose ; isomerization ; xylulose ; biotechnology ; Bacillus (bacteria)
Language	English
Dates of publication	1994-12
Size	p. 4495-4499.
Document type	Article
ZDB-ID	223011-2
ISSN	1098-5336 ; 0099-2240
ISSN (online)	1098-5336
ISSN	0099-2240
Database	NAL-Catalogue (AGRICOLA)

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Article: Sequence of a cDNA encoding the ferritin H-chain from an 11-week-old human fetal brain.

Dhar, M / Chauthaiwale, V / Joshi, J G

Gene

1993 Volume 126, Issue 2, Page(s) 275–278

Abstract: A cDNA library in lambda Charon BS(-) from 11-week-old human fetal brain (FB) was screened using a human liver ferritin (FTH)-encoding cDNA as a probe. The complete sequence of the positive clone, cFB1, showed that the coding region and a part of the 5' ... ...

Abstract	A cDNA library in lambda Charon BS(-) from 11-week-old human fetal brain (FB) was screened using a human liver ferritin (FTH)-encoding cDNA as a probe. The complete sequence of the positive clone, cFB1, showed that the coding region and a part of the 5' and 3' untranslated regions (UTR) are identical to the corresponding published sequence of the liver cDNA. However, a particularly noteworthy difference is the presence of 279 bp of additional sequence in the FB 3'-UTR. Northern blot analysis of FB poly(A)+RNA showed it to be a part of the FTH transcript. Comparison of the 279-bp sequence with the GenBank and EMBL databases showed it to be 94.1, 62.5, and 58.9% similar to segments from human, mouse and rat FTH genomic sequences, respectively. However, in all these cases, only a part of this 279-bp sequence has been found in the nontranscribed region. We therefore conclude that in FB, the 279-bp sequence is a part of the mature FTH mRNA. Sequence analysis also suggests a differential poly(A) site selection in the production of FTH mRNA in FB and liver.
MeSH term(s)	Amino Acid Sequence ; Base Sequence ; Brain/embryology ; Brain/metabolism ; Cloning, Molecular ; DNA ; Ferritins/genetics ; Fetus ; Humans ; Molecular Sequence Data ; Sequence Homology, Nucleic Acid ; Transcription, Genetic
Chemical Substances	DNA (9007-49-2) ; Ferritins (9007-73-2)
Language	English
Publishing date	1993-04-30
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	391792-7
ISSN	1879-0038 ; 0378-1119
ISSN (online)	1879-0038
ISSN	0378-1119
DOI	10.1016/0378-1119(93)90380-l
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Zs.A 1357: Show issues

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Article: Bacteriophage lambda as a cloning vector.

Chauthaiwale, V M / Therwath, A / Deshpande, V V

Microbiological reviews

1992 Volume 56, Issue 4, Page(s) 577–591

Abstract: Extensive research has been directed toward the development of multipurpose lambda vectors for cloning ever since the potential of using coliphage lambda as a cloning vector was recognized in the late 1970s. An understanding of the intrinsic molecular ... ...

Abstract	Extensive research has been directed toward the development of multipurpose lambda vectors for cloning ever since the potential of using coliphage lambda as a cloning vector was recognized in the late 1970s. An understanding of the intrinsic molecular organization and of the genetic events which determine lysis or lysogeny in lambda has allowed investigators to modify it to suit the specific requirements of gene manipulations. Unwanted restriction sites have been altered and arranged together into suitable polylinkers. The development of a highly efficient in vitro packaging system has permitted the introduction of chimeric molecules into hosts. Biological containment of recombinants has been achieved by introducing amber mutations into the lambda genome and by using specific amber suppressor hosts. Taking advantage of the limited range of genome size (78 to 105% of the wild-type size) for its efficient packaging, an array of vectors has been devised to accommodate inserts of a wide size range, the limit being 24 kbp in Charon 40. The central dispensable fragment of the lambda genome can be replaced by a fragment of heterologous DNA, leading to the construction of replacement vectors such as Charon and EMBL. Alternatively, small DNA fragments can be inserted without removing the dispensable region of the lambda genome, as in lambda gt10 and lambda gt11 vectors. In addition, the introduction of many other desirable properties, such as NotI and SfiI sites in polylinkers (e.g., lambda gt22), T7 and T3 promoters for the in vitro transcription (e.g., lambda DASH), and the mechanism for in vivo excision of the intact insert (e.g., lambda ZAP), has facilitated both cloning and subsequent analysis. In most cases, the recombinants can be differentiated from the parental phages by their altered phenotype. Libraries constructed in lambda vectors are screened easily with antibody or nucleic acid probes since several thousand clones can be plated on a single petri dish. Besides the availability of a wide range of lambda vectors, many related techniques such as rapid isolation of lambda DNA, a high efficiency of commercially available in vitro packaging extracts, and in vitro amplification of DNA via the polymerase chain reaction have collectively contributed to lambda's becoming one of the most powerful and popular tools for molecular cloning.
MeSH term(s)	Bacteriophage lambda/genetics ; Bacteriophage lambda/growth & development ; Cloning, Molecular/methods ; Escherichia coli/genetics ; Genetic Vectors/genetics ; Recombination, Genetic
Language	English
Publishing date	1992-12
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	6864-0
ISSN	0146-0749
ISSN	0146-0749
DOI	10.1128/mr.56.4.577-591.1992
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Article: pRb and p107 regulate E2F activity during lens fiber cell differentiation.

Rampalli, A M / Gao, C Y / Chauthaiwale, V M / Zelenka, P S

Oncogene

1998 Volume 16, Issue 3, Page(s) 399–408

Abstract: During growth arrest and differentiation, activity of the E2F family of transcription factors is inhibited by interactions with pRb and the related proteins, p107 and p130. To determine which members of the E2F and pRb families may contribute to growth ... ...

Abstract	During growth arrest and differentiation, activity of the E2F family of transcription factors is inhibited by interactions with pRb and the related proteins, p107 and p130. To determine which members of the E2F and pRb families may contribute to growth arrest as lens epithelial cells differentiate into fiber cells, we examined the expression of individual E2F species and characterized the E2F protein complexes formed in rat lens epithelia and fibers. RT/PCR detected all five known members of the E2F family in lens epithelial cells, but only E2F-1, E2F-3, and E2F-5 in fiber cells. Proteins extracted from lens epithelia of newborn rats formed at least two specific complexes with an E2F consensus oligonucleotide. Proteins from lens fiber cells formed three specific complexes, one of which comigrated with an epithelial cell complex. Incubation of epithelial and fiber cell extracts with an antibody specific for p107 demonstrated that two fiber cell complexes and one epithelial cell complex contained p107. Although the remaining fiber cell complex did not react with antibodies to pRb or p130 in this assay, a strong reaction with pRb antibody was observed when the electromobility shifted complexes were subsequently immunoblotted (shift/Western assay). Immunocytochemistry confirmed that pRb protein is present in the nuclei of both epithelial cells and fiber cells. Immunoblotting of whole cell extracts with pRb antibody showed multiple, phosphorylated forms of pRb in the epithelial cells, but predominantly hypophosphorylated pRb in the fiber cells. None of the complexes formed with E2F were recognized exclusively by the p130 antibody, although the previously identified p107 complexes reacted weakly. The absence of p130/E2F complexes was correlated with the presence of multiple ubiquitinated forms of p130, especially in the fiber cells. Thus, although p130/E2F complexes are implicated in the terminal differentiation of many cell types, in differentiating lens fiber cells pRb and p107 seem to be the primary regulators of E2F activity.
MeSH term(s)	Animals ; Animals, Newborn ; Carrier Proteins ; Cell Cycle Proteins ; Cell Differentiation ; DNA/metabolism ; DNA-Binding Proteins ; E2F Transcription Factors ; E2F1 Transcription Factor ; E2F3 Transcription Factor ; E2F5 Transcription Factor ; Immunoblotting ; Immunoenzyme Techniques ; Lens, Crystalline/cytology ; Lens, Crystalline/metabolism ; Nuclear Proteins/biosynthesis ; Nuclear Proteins/metabolism ; Phosphoproteins/biosynthesis ; Phosphoproteins/metabolism ; Proteins ; Rats ; Rats, Wistar ; Retinoblastoma Protein/biosynthesis ; Retinoblastoma Protein/metabolism ; Retinoblastoma-Binding Protein 1 ; Retinoblastoma-Like Protein p130 ; Transcription Factor DP1 ; Transcription Factors/biosynthesis ; Transcription Factors/genetics ; Transcription Factors/metabolism ; Ubiquitins/metabolism
Chemical Substances	Carrier Proteins ; Cell Cycle Proteins ; DNA-Binding Proteins ; E2F Transcription Factors ; E2F1 Transcription Factor ; E2F3 Transcription Factor ; E2F5 Transcription Factor ; E2f1 protein, rat ; Nuclear Proteins ; Phosphoproteins ; Proteins ; Rbl2 protein, rat ; Retinoblastoma Protein ; Retinoblastoma-Binding Protein 1 ; Retinoblastoma-Like Protein p130 ; Transcription Factor DP1 ; Transcription Factors ; Ubiquitins ; DNA (9007-49-2)
Language	English
Publishing date	1998-01-22
Publishing country	England
Document type	Journal Article
ZDB-ID	639046-8
ISSN	1476-5594 ; 0950-9232
ISSN (online)	1476-5594
ISSN	0950-9232
DOI	10.1038/sj.onc.1201546
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Article: Genetic transformation of Chainia and heat attenuation of its restriction system.

Chauthaiwale, V M / Vyas, P R / Deshpande, V V

Canadian journal of microbiology

1991 Volume 37, Issue 9, Page(s) 713–715

Abstract: A PEG-mediated transformation system for Chainia (NCL 82-5-1) was developed using a broad host range Streptomyces vector, pIJ702. Protoplasts prepared from Chainia (NCL 82-5-1) were regenerated with 5% efficiency. Transformation of the protoplasts with ... ...

Abstract	A PEG-mediated transformation system for Chainia (NCL 82-5-1) was developed using a broad host range Streptomyces vector, pIJ702. Protoplasts prepared from Chainia (NCL 82-5-1) were regenerated with 5% efficiency. Transformation of the protoplasts with pIJ702 gave 10-20 transformants/micrograms DNA. The low efficiency of transformation is attributed to a restriction system in Chainia; this could be inhibited by treating the protoplasts at 42 degrees C for 10 min just before transformation. The yield of transformants increased 100-fold when pIJ702 was modified by passage in Chainia. Because the plasmid replicon was functional in Chainia and the modified plasmid was stably maintained, the transformation system should be useful for self-cloning in Chainia NCL 82-5-1 of the many commercially important enzymes this strain is known to produce.
MeSH term(s)	Cloning, Molecular/methods ; DNA Restriction Enzymes/genetics ; Genetic Vectors/genetics ; Hot Temperature ; Plasmids/genetics ; Polyethylene Glycols ; Protoplasts/metabolism ; Streptomyces/genetics ; Temperature ; Transformation, Bacterial/genetics
Chemical Substances	Polyethylene Glycols (3WJQ0SDW1A) ; DNA Restriction Enzymes (EC 3.1.21.-)
Language	English
Publishing date	1991-09
Publishing country	Canada
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	280534-0
ISSN	1480-3275 ; 0008-4166
ISSN (online)	1480-3275
ISSN	0008-4166
DOI	10.1139/m91-121
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Zs.B 85: Show issues

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Article: A novel ferritin heavy chain messenger ribonucleic acid in the human brain.

Joshi, J G / Fleming, J T / Dhar, M / Chauthaiwale, V

Journal of the neurological sciences

1995 Volume 134 Suppl, Page(s) 52–56

Abstract: In the aging human brain, the concentrations of iron and its major storage protein, ferritin, rise but the distribution of metal and protein remains non-uniform. More ferritin could be isolated from the brains of humans who died of Alzheimer's disease ( ... ...

Abstract	In the aging human brain, the concentrations of iron and its major storage protein, ferritin, rise but the distribution of metal and protein remains non-uniform. More ferritin could be isolated from the brains of humans who died of Alzheimer's disease (AD) than from age- and sex-matched controls. Also, brain ferritin of rats chronically exposed to aluminum chloride in their drinking water contained more aluminum and iron. Based on these earlier observations, a more detailed study of human brain ferritin was initiated. The results showed that ferritin is a component of neuritic (senile) plaques in AD. Ferritin obtained from normal or AD brains is composed of 24 subunits (70% heavy (H) chain; 30% light (L) chain). With high performance liquid chromatography, the subunits resolved into a cluster of four H-chain peaks and one major L-chain peak. Western blot analysis confirmed the identity of H- and L-fractions. The techniques of molecular biology revealed the presence of an additional ferritin messenger ribonucleic acid (mRNA) species for the H subunit which was more abundant in the brain than in other human tissues. It contained the entire sequence of 919 nucleotides of H chain mRNA from liver but also an additional segment of 279 nucleotides in the 3'-untranslated region. The two mRNA seemed to arise by the use of an alternate polyadenylation site of the same primary transcript. Ribonuclease protection assays revealed that the concentrations of the longer mRNA in the normal hippocampus and the hippocampus of patients with AD brains were similar.
MeSH term(s)	Aluminum/metabolism ; Alzheimer Disease/metabolism ; Blotting, Northern ; Blotting, Western ; Brain Chemistry/physiology ; Chromatography, High Pressure Liquid ; Ferritins/biosynthesis ; Hippocampus/metabolism ; Humans ; Iron/metabolism ; RNA, Messenger/biosynthesis
Chemical Substances	RNA, Messenger ; Ferritins (9007-73-2) ; Aluminum (CPD4NFA903) ; Iron (E1UOL152H7)
Language	English
Publishing date	1995-12
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80160-4
ISSN	1878-5883 ; 0022-510X ; 0374-8642
ISSN (online)	1878-5883
ISSN	0022-510X ; 0374-8642
DOI	10.1016/0022-510x(95)00208-j
Database	MEDical Literature Analysis and Retrieval System OnLINE

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