Megadeth's Countdown to Extinction PDF

By Megadeth

ISBN-10: 0793536669

ISBN-13: 9780793536665

Eleven songs, together with: dermis O' My tooth * Symphony Of Destruction * Sweating Bullets * Countdown To Extinction * and extra.

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02 M ammonium formate for 24 h. The ammonium formate used to prepare the eluant was freshly sublimed. 02 M ammonium formate and applied to the column that was then eluted with the same salt solution at a flow rate of II ml/h. Fractions of 3 ml were collected. Small portions were removed for bioassay and the active portion of the eluate between 228 and 255 ml was lyophilized to yield 206 JLg proctolin (determined by bioassay). With Sephadex G 15, elution volumes of I-JLmol quantities of (a) glycylleucyl-tyrosine, (b) leucyl-tyrosine, and (c) tyrosine amide occurred in the expected order, a < b < c.

After elution of the basic fraction, the column was regenerated by washing with 3 vol. each 2 N sodium hydroxide, water, and 2 N hydrochloric acid, followed by water until the washings were neutral. By using a rotary evaporator, the solvent was removed at reduced pressure (water aspirator) and room temperature from the 4 N ammonium hydroxide-ethanol eluate, which contained proctolin. The residue (approximately 30 g from the extract of 25000 cockroaches) was dissolved in 2 liters water and was applied to a 3 x 80 cm column of Dowex 50W-x8 (50 to 100 mesh, NH4+ form).

Chromatography of the dansyl derivative showed a spot that could be attributed to threonine, the fifth amino acid detected in the hydrolysate of proctolin. The position of this spot was different from that of Dns-threonine amide, indicating that proctolin was not an amide. As a result of these studies, proctolin was proposed to be H-Arg-Tyr-Leu-Pro-Thr-OH. C. Synthesis of Proctolin and Comparison of Synthetic and Natural Proctolin The peptide with the sequence proposed for proctolin was synthesized by both the mixed anhydride procedure and the pentaftuorophenyl ester method.

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Countdown to Extinction by Megadeth

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