Fractionatton of Brain Macromolecules—ii. Isolation of Protein‐linked Sialomucopoly‐saccharides From Subcellular, Particulate Fractions From Rat Brain

Brunngraber, Eric G.; Brown, Barbara D.

doi:10.1111/j.1471-4159.1964.tb11604.x

Cited by 49 publications

(9 citation statements)

References 24 publications

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“…crude mitochondrial, microsomal and nuclear pellets (Brunngraber & Brown, 1964) was similar to that of the gangliosides. To draw appropriate conclusions with regard to their subcellular localization, it was necessary to prepare subcellular fractions of a higher degree of purity than those previously studied.…”

mentioning

confidence: 57%

“…Preparation of 8ubcellular fradions. The crude mitochondrial and microsomal pellets from three rat whole brains were obtained as described by Brunngraber & Brown (1964), except that the microsomal pellet was sedimented by centrifuging for 90min. at 105400g in the no.…”

Section: Methodsmentioning

confidence: 99%

“…Osmotic-'shock' treatment of the crude mitochondrial pellet from whole rat brain consisted of successively suspending the mitochondrial pellet obtained by procedures described by Brunngraber & Brown (1964) in 0-02M-trischloride buffer, pH8-0, centrifuging at 105000g for 30min., resuspending in water and recentrifuging. Suspension of the particulate fraction was performed by homogenizing by hand in a TenBroeck homogenizer.…”

Section: Methodsmentioning

confidence: 99%

“…The present work is a continuation of investigations designed to ascertain the suboellular localization of the insoluble glycoproteins associated with particulate fractions obtained from rat whole brain. Brunngraber & Brown (1964, 1967 reported that most of the protein-bound NANA* of the isolated particulate matter of rat brain after the gangliosides are extracted is present in a proteinlinked sialomucopolysaccharide. The sialomucopolysaccharides contain NANA, hexose, hexosamine and fucose; the sialomucopolysaccharide molecule consists of a repeating unit containing hexosamine and hexose to which NANA and fucose are attached.…”

mentioning

confidence: 99%

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The distribution of protein-bound N-acetylneuraminic acid in subcellular fractions of rat brain

Brunngraber¹,

Dekirmenjian²,

Brown³

1967

Biochemical Journal

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Protein-bound N-acetylneuraminic acid and hexosamine, including the sialomucopolysaccharides, occur mainly in the least dense particles sedimented in the microsomal fraction from rat whole brain. Particles rich in protein-bound N-acetylneuraminic acid and hexosamine are also found in the subcellular fraction separated as a layer between 0.8m- and 1.2m-sucrose after centrifuging the crude mitochondrial preparation in a density gradient. This distribution is similar to that of the gangliosides and suggests an association of all of these substances in the same subcellular structures. It is postulated that the sialomucopolysaccharides, as well as the gangliosides, are components of cell membranes. Evidence is presented that indicates that there are quantitative differences between distribution of the gangliosides on the one hand, and protein-bound N-acetylneuraminic acid and hexosamine on the other. The ratio of protein-bound N-acetylneuraminic acid (and hexosamine) to gangliosidic N-acetylneuraminic acid (and hexosamine) present in individual subcellular fractions obtained by density-gradient centrifugation tends to increase with increasing particle density. Exposure of the crude mitochondrial fraction to osmotic ;shock' before density-gradient centrifugation causes a shift of the protein-bound N-acetylneuraminic acid and gangliosides to the less dense fractions. In some experiments, a selective shift of the protein-bound N-acetylneuraminic acid was observed.

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mentioning

confidence: 57%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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The distribution of protein-bound N-acetylneuraminic acid in subcellular fractions of rat brain

Brunngraber¹,

Dekirmenjian²,

Brown³

1967

Biochemical Journal

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“…The glycopeptides were subsequently digested with carboxypeptidase A-DFP and Ieucine a~no~ptidase according to Li et al [7]. Nucleic acids and mucopolysaccharides were eliminated with cetyl pyridinium chloride as described by Brunngraber et al [8]. The glycopeptides were then separated from most of the contaminating peptides and amino acids by gel filtration in ~sti~ed water on Sephadex G-l 5.…”

Section: Preparation Of Glycopep Tidesmentioning

confidence: 99%

The composition of glycopeptides, derived from neural membranes, which affect neurite growth in vitro

et al. 1972

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Role of glycoproteins in the uptake of L‐ ¹⁴C ‐ leucine by a synaptosomal fraction

Rossowska

1980

J of Neuroscience Research

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The present study deals with the effect of modification of membrane glycoproteins on the accumulation of amino acids by isolated synaptosomal fractions, measured by the uptake of 14C-leucine. Superficial sugar receptors were modified by concanavalin A, neuraminidase, and neuraminidase followed by concanavalin A binding. It is demonstrated that neuraminidase treatment followed by concanavalin A binding results in significant diminution of 14C-leucine uptake. A similar effect was observed in experimental hypox followed by concanavalin A binding to isolated synaptosomal fractions. In this case, however, (Na+-K+) -ATPase activity remained unchanged. These results seem to indicate that 14C-leucine uptake was dependent on integrity of glycoprotein structure of synaptosomal membranes.

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Fractionatton of Brain Macromolecules—ii. Isolation of Protein‐linked Sialomucopoly‐saccharides From Subcellular, Particulate Fractions From Rat Brain

Cited by 49 publications

References 24 publications

The distribution of protein-bound N-acetylneuraminic acid in subcellular fractions of rat brain

The distribution of protein-bound N-acetylneuraminic acid in subcellular fractions of rat brain

The composition of glycopeptides, derived from neural membranes, which affect neurite growth in vitro

Role of glycoproteins in the uptake of L‐ ¹⁴C ‐ leucine by a synaptosomal fraction

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Fractionatton of Brain Macromolecules—ii. Isolation of Protein‐linked Sialomucopoly‐saccharides From Subcellular, Particulate Fractions From Rat Brain

Cited by 49 publications

References 24 publications

The distribution of protein-bound N-acetylneuraminic acid in subcellular fractions of rat brain

The distribution of protein-bound N-acetylneuraminic acid in subcellular fractions of rat brain

The composition of glycopeptides, derived from neural membranes, which affect neurite growth in vitro

Role of glycoproteins in the uptake of L‐ 14C ‐ leucine by a synaptosomal fraction

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Role of glycoproteins in the uptake of L‐ ¹⁴C ‐ leucine by a synaptosomal fraction