Thin-layer chromatography of 1-dimethylaminonaphthalene-5-sulphonyl derivatives of amino acids present in superfusates of cat cerebral cortex

Crowshaw, K.; Jessup, Sheila J.; Ramwell, Peter W.

doi:10.1042/bj1030079

Cited by 140 publications

(32 citation statements)

References 11 publications

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“…This may have some functional significance, since the concentration of GABA in the plasma has been estimated at 2-4 gM in cats (Crowshaw, Jessup & Ramwell, 1967).…”

Section: Discussionmentioning

confidence: 99%

Influence of Neuroglial Transport on the Action of Γ‐aminobutyric Acid on Mammalian Ganglion Cells

Brown

Galvan

1977

British J Pharmacology

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1The effect of inhibiting the transport of γ‐aminobutyric acid (GABA) by neuroglial cells on the depolarizing action of exogenous amino acids on isolated superior cervical ganglia of the rat was studied. 2Transport (measured by uptake of [3H]‐GABA) was inhibited by (a) reducing external [Na+] from 143 to 2 mm and (b) administering alternative carrier‐substrates, 3‐amino‐n‐butyric acid (β‐amino‐butyric acid, BABA) and (±)‐nipecotic acid at a concentration of 1 mm. 3All three procedures enhanced the depolarization produced by low concentrations of GABA (<10 μm) but did not alter the maximum response, nor the response to 3‐aminopropanesulphonic acid (a gabamimetic with low affinity for the neuroglial carrier). 4It is concluded that the neuroglial uptake process can limit the action of exogenous GABA upon neurones, by reducing the interstitial GABA concentration.

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“…This may have some functional significance, since the concentration of GABA in the plasma has been estimated at 2-4 gM in cats (Crowshaw, Jessup & Ramwell, 1967).…”

Section: Discussionmentioning

confidence: 99%

Influence of Neuroglial Transport on the Action of Γ‐aminobutyric Acid on Mammalian Ganglion Cells

Brown

Galvan

1977

British J Pharmacology

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“…A two-dimensional development using water-formic acid (100 : 1.5) and benzene-acetic acid (9: l), respectively [15], was first carried out and the plates inspected. In the same direction as the second dimension a third run was then performed using ethyl acetate-methanolacetic acid (20: 1 : 1) [16] to distinguish Dan-Ser/DanThr, Dan-Asp/Dan-Glu, Dan-Ala/Dan-NH, and DanCys(Cm)/Dan-OH, and finally a fourth run using one part of ethanol to three parts of pyridine-acetic acid-water (9: 16: 1000, pH 4.4) to separate dansylderivatives of the basic amino acids [17].…”

Section: Methodsmentioning

confidence: 99%

Horse Liver Alcohol Dehydrogenase

Jörnvall

1970

European Journal of Biochemistry

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1. The ethanol-active isoenzyme of horse liver alcohol dehydrogenase has been carboxymethylated with iodo[14C]acetate and different samples of the labelled protein treated with trypsin (both with and without previous maleylation of the substrate), chymotrypsin, pepsin or cyanogen bromide. In each case most of the resultant, peptides have been purified and characterized.2. Data are given for those peptides originating from an N-terminal region comprising about 4001, of the protein chain and the amino acid sequence of the first 140 residues is deduced.3. It is established that over this region the two chains of the enzyme have an identical sequence. The N-terminus is an acetylated serine residue and the "active site" cysteine residue occupies position no. 46. One oft he two tryptophans of the chain, but none of the four tyrosines, is in the sequence given. The region around position 100 is rich in cysteine residues. Some serine residues were found to have reacted during maleylation of the protein. 4.The amino acid sequence is compared to the previously known sequences of glyceraldehyde 3-phosphate dehydrogenase from two species. Similarities strongly suggest a true homology, a t least of an N-terminal region, and support theories of a common ancestral origin for these enzymes. The reactive cysteine residues are, however, a t widely different positions in the primary structures.

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“…Dansyl amino acids were identified by chromatography on polyamide thin-layer sheets (7.5 x 7.5 cm) [19]. Additional solvent systems were used to distinguish between the dansyl derivatives of hydroxy amino acids and of acidic amino acids [20] and between the monodansyl derivatives of histidine, lysine and arginine [21].…”

Section: N-terminal Amino-acid Analysismentioning

confidence: 99%

The Amino-Acid Sequence and Carbohydrate Content of Phospholipase A2 from Bee Venom

et al. 1974

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The complete amino acid sequence of phospholipase A2 from the venom of the common European honey-bee (Apis mellifica) has been determined. The sequence of amino acid residues at the N-terminus was obtained by direct application of the Edman degradation technique and that at the C-terminus by digestion with carboxypeptidasesA and B. Digestion of the reduced and carboxymethylated enzyme with trypsin yielded a completely soluble peptide mixture, all the components of which were isolated and their structures determined. Overlaps of the tryptic peptides were deduced from the structures of peptides isolated after digestion of the reduced and carboxymethylated enzyme with chymotrypsin and of the reduced and aminoethylated enzyme with a protease specific for cleavage at lysine residues. Two remaining ambiguities were resolved by peptides obtained from a digest of the reduced, carboxymethylated and maleylated enzyme with trypsin. The phospholipase A2 consists of a single chain of 128 amino acid residues and contains attached carbohydrate residues. The composition and point of attachment of the carbohydrate moiety have been established. Phospholipase A2 is the name given to a family of enzymes which catalyse the hydrolysis of fatty acid ester bonds at the 2-carbon position of 3-sn-phosphoglycerides. Enzymes with this specificity have been isolated from pancreatic tissue [l], bee venom [2] and a variety of snake venoms [3]. The enzymes have certain characteristics in common, for example relatively small size, high disulphide-bridge content, heat stability and requirement for Ca2+ ions for activity. There are, however, important differences. In particular, the pancreatic enzyme appears to be active on the aggregated (micellar) form of phospholipids [4], whereas the substrate for the bee-venom enzyme has been shown to be the dispersed (monomolecular) phospholipid [2]. It is also of interest that the enzyme from pancreas is secreted as an inactive zymogen which may be activated by limited proteolysis with trypsin sequence of the zymogen and the positions of the disulphide bridges in porcine pancreatic phospholipase A2 have been reported [6,7].Our interest in phospholipase A2 from bee venom arose from the observation [2] that the enzyme crystallises readily which, combined with its relatively small size, suggests that it may be amenable to total structure determination. As a preliminary to crystallographic analysis we have determined the primary structure of the enzyme; this work has been the subject of a preliminary report [S]. A missing overlap in the earlier work has now been established with certainty and three corrections have been made to the previously published sequence. In addition, the composition and point of attachment of the carbohydrate moiety have been established. An interesting aspects of the present work has been the use of a recently characterized protease isolated from the basidiomycete Armillaria mellea [9] ; the enzyme has been shown to have specificity for cleavage of peptide bonds N-terminal to lysin...

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Thin-layer chromatography of 1-dimethylaminonaphthalene-5-sulphonyl derivatives of amino acids present in superfusates of cat cerebral cortex

Cited by 140 publications

References 11 publications

Influence of Neuroglial Transport on the Action of Γ‐aminobutyric Acid on Mammalian Ganglion Cells

Influence of Neuroglial Transport on the Action of Γ‐aminobutyric Acid on Mammalian Ganglion Cells

Horse Liver Alcohol Dehydrogenase

The Amino-Acid Sequence and Carbohydrate Content of Phospholipase A2 from Bee Venom

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