Zwei Universalreagentien zur Dünnschichtchromatographie

Ertel, H.; Horner, L.

doi:10.1016/s0021-9673(01)86410-4

Cited by 86 publications

(7 citation statements)

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“…The amino acid derivatives and peptides were located by spraying the chromatograms with the ninhydrin reagent 1 10 1 for free amino groups, the hydrazide reagent' 11 1 for hydrazides and the modified chlorine reagent' 12 J for all peptide derivatives. The amino acid derivatives used were prepared according to known methods^1 3 '.…”

Section: Methodsmentioning

confidence: 99%

Synthetic and Binding Studies on the Postulated Calcium Binding Site I of Calmodulin

Marchiori¹,

Borin²,

Chessa³

et al. 1983

Hoppe-Seyler´s Zeitschrift für physiologische Chemie

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The synthesis of the dodecapeptide [sequence (20-31)] representing the hypothetical calcium binding site I of calmodulin by classical methods in solution is described. The interaction of this synthetic calmodulin fragment with calcium ions has been investigated by CD in water and in 98% trifluoroethanol solution. Calcium ions have no effect on the dichroic absorption of the dodecapeptide in aqueous solution in the wavelength range 200-250 nm. However, in 98% trifluoroethanol a linear variation of the CD properties has been observed as a function of the Ca 2e /peptide molar ratio. The CD results indicate the formation of a 1:1 calcium-dodecapeptide complex with a very high binding constant. Synthetische und Bindungsuntersuchungen über die postulierte Calcium-Bindungsstelle I des CalmodulinsZusammenfassung: Es wird die Synthese des linearen Dodecapeptids der Sequenz 20-31 des Calmodulins als mögliche Calcium-Bindungsstelle beschrieben. Mittels Circulardichroismus wurde die Wechselwirkung zwischen diesem synthetischen Calmodulin-Fragment mit Calcium-Ionen in Wasser und 98% Trifluorethanol untersucht. In Wasser wurde der Dichroismus des Dodecapeptids im Wellenlängenbereich 200-250 nm nicht beeinflußt, während in 98% Trifluorethanol eine lineare Abhängigkeit des Circulardichroismus als Funktion des molaren Verhältnisses Ca 20 /Peptid gemessen wurde. Die Resultate der spektroskopischen Untersuchungen sprechen für einen 1:1-Calcium/Dodecapeptid-Komplex mit hoher Bindungskonstante.

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Section: Methodsmentioning

confidence: 99%

Synthetic and Binding Studies on the Postulated Calcium Binding Site I of Calmodulin

Marchiori¹,

Borin²,

Chessa³

et al. 1983

Hoppe-Seyler´s Zeitschrift für physiologische Chemie

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“…The plates were developed with the solvent system of n-propanol-ammonia-water, 6:1:3 (vol/vol/vol) (5). The components were located by spraying with aniline-phthalate (Brinkmann) and potassium permanganate-sulfuric acid (6).…”

Section: Methodsmentioning

confidence: 99%

Selective adsorption of heterophile polyglycerophosphate antigen from antigen extracts of Streptococcus mutans and other gram-positive bacteria

Hamada¹,

Tai²,

Slade³

1976

Infect Immun

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Hot saline extracts of Streptococcus mutans have been shown to contain antigenic substances which occasionally react nonspecifically with some antisera against whole cells of various serological groups and types of streptococci. Chromatography of the extract of S. mutans strain MT703 (serotype e) on a diethylaminoethyl-Sephadex A-25 column gave two principal antigens. One antigen was eluted without adsorption to the resin and was identified as the serotype-specific polysaccharide. The other antigen, which contained a large quantity of phosphorus, was adsorbed to and released from the resin by gradient elution. It was reactive against the antisera specific for polyglycerophosphate (PGP) from group A Streptococcus pyogenes and/or S. mutans strain Ingbritt (type c). The PGP antigen was further purified by gel filtration with Sephadex G-75. Two peaks, PGP-1 and PGP-2, were obtained. Each possessed the same antigenic specificity to anti-PGP serum as shown by immunodiffusion. Chemical analyses revealed that the molar ratio of phosphorus to glycerol in both was about 1:1, although the protein content between the two was significantly different. PGP antigen was found to be widely distributed in hot saline extracts from various gram-positive bacteria, with a few exceptions. However, all gramnegative bacteria examined were free of PGP. The PGP in the hot saline extracts of various gram-positive bacteria possessed an essentially identical antigenic specificity. The addition of diethylaminoethyl-Sephadex A-25 resin to hot saline extracts successfully removed the cross-reacting PGP antigen. After adsorption of the extract from S. mutans, the supernatant contained only type-specific polysaccharide antigen, except type b, in which both type b-specific polysaccharide and PGP antigens were adsorbed with the resin. This simple procedure should be useful for the removal of the PGP-type teichoic acid from antigen extracts of bacteria that contain uncharged polysaccharides. Umemoto (New York State University School of Dentistry at Buffalo, Buffalo), B. Guggenheim 903 on July 15, 2020 by guest http://iai.asm.org/ Downloaded from

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“…Optical rotations were determined with a Perkin-Elmer 141 polarimeter. Samples for elerncntal analyses wcre dried in ~'ucuo over P205 at 4 0 ' C Thin-layer chromatography on silica gel plates F-254 (Merck AG, Darmstadt) were carried out in the following systems : (1) Amino acid derivatives and peptides were located by spraying the chromatograms with the ninhydrin reagent [8] for free amino groups, the hydrazide reagent [9] for hydrazides and the modified chlorine reagent [lo] for all peptide derivatives.…”

Section: Twhnical Details O F P C~p Tide S J W Hes Ismentioning

confidence: 99%

“…Amino acid derivatives and peptides were located by spraying the chromatograms with the ninhydrin reagent [8] for free amino groups, the hydrazide reagent [9] for hydrazides and the modified chlorine reagent [lo] for all peptide derivatives.…”

Section: Twhnical Details O F P C~p Tide S J W Hes Ismentioning

confidence: 99%

Synthetic peptides reproducing the site phosphorylated by cAMP‐dependent protein kinase in protein phosphatase inhibitor‐1

Chessa

Borin

Marchiori

et al. 1983

European Journal of Biochemistry

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The hexapeptide (Arg),-Pro-Thr-Pro-Ala (T 1) and octapeptide (Arg),-Pro-Thr-Pro-Ala (T5), reproducing the phosphorylatable site of protein phosphatase inhibitor-I , a physiological target of CAMP-dependent protein kinase, and five related peptides were synthesized by the method in solution.The phosphorylation rates of such peptides by CAMP-dependent protein kinase and their kinetic parameters have been determined and compared with those of the hexapeptide (Arg),-Ah-Ser-Val-Ah, reproducing the phosphorylatable site of rat liver pyruvate kinase. The results obtained show that both the presence of threonine instead of serine and the adjacent C-terminal proline represent highly unfavourable factors seriously impairing the protein kinase reaction by both increasing K,, and depressing V. On the other hand the N-terminal proline is compatible with high phosphorylation rates and the row of four rather than two consecutive arginines improves the phosphorylation efficiency by lowering tenfold the K,,, without affecting the V. The extension of the hexapeptide T 1 on its C-terminal side to give the derivative (Arg),-Pro-Thr-Pro-Ala-Thr-Val-AIa has no significant effect on the kinetic parameters.Moreover no relationship between the phosphorylation efficiency and the predicted secondary structures around the target residue could be evidenced.Therefore the local structural features of the phosphorylatable site of inhibitor-I cannot fully account for the fast phosphorylation of this regulatory protein. Other factors must optimize the protein kinase reaction.CAMP-dependent protein kinase is a multi-substrate enzyme capable of modulating the biological activity of a variety of proteins by catalyzing the phosphorylation of specific amino acid residues (for a review see [I]). In most instances the phosphorylated residues arc serines. However in the case of protein phosphatase inhibitor-I a threonyl residue is modified by CAMP-dependent protein kinase [2].The factors influencing the phosphorylation of seryl residues by CAMP-dependent protein kinase have been thoroughly studied with the aid of synthetic peptides which corrcspond to the amino acid sequences around the phosphorylation sites in protein substrates [3 -61. All these reports agree on the requirement ofa pair of adjacent basic amino acid residues either Arg-Arg or Arg-Lys close to the N-terminal side ofthe phosphorylated serine. On the other hand the length of the peptide, beyond 6-7 residues, is not so important; actually small peptides are often better substrates than the intact proteins, indicating that the integrity of the protein substrate is by no means required for phosphorylation [l].A structural modification which dramatically impairs the phosphorylation efficiency of these peptides is the replacement of threonine for serine, giving derivatives with both higher K,,, and lower Vvalues [3,4,6] per se. The sequence surrounding Thr-35 is unusual for two more reasons: the threonine is flanked by two prolyl residues and there are four rather than two arginines [2].In ord...

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Zwei Universalreagentien zur Dünnschichtchromatographie

Cited by 86 publications

References 1 publication

Synthetic and Binding Studies on the Postulated Calcium Binding Site I of Calmodulin

Synthetic and Binding Studies on the Postulated Calcium Binding Site I of Calmodulin

Selective adsorption of heterophile polyglycerophosphate antigen from antigen extracts of Streptococcus mutans and other gram-positive bacteria

Synthetic peptides reproducing the site phosphorylated by cAMP‐dependent protein kinase in protein phosphatase inhibitor‐1

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