Improved Method for Hydrolyzing Proteins and Peptides Without Inducing Racemization and for Determining Their True D-Amino Acid Content

D’Aniello, Antimo; Petrucelli, L.; Gardner, C. E.; Fisher, G. H.

doi:10.1006/abio.1993.1423

Cited by 28 publications

(13 citation statements)

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“…The enzymatic procedure was performed according to the method described by D'Aniello et al (19) and modified as follows. In three Eppendorf tubes (marked 'sample', 'sample plus internal standard', and 'blank sample') was added 20 l of the purified sample.…”

Section: Methods 1: Enzymatic Fluorometricmentioning

confidence: 99%

Occurrence of D‐aspartic acid and N‐methyl‐D‐aspartic acid in rat neuroendocrine tissues and their role in the modulation of luteinizing hormone and growth hormone release

D‧Aniello¹,

et al. 2000

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Using two specific and sensitive fluorometric/HPLC methods and a GC-MS method, alone and in combination with D-aspartate oxidase, we have demonstrated for the first time that N-methyl-D-aspartate (NMDA), in addition to D-aspartate (D-Asp), is endogenously present as a natural molecule in rat nervous system and endocrine glands. Both of these amino acids are mostly concentrated at nmol/g levels in the adenohypophysis, hypothalamus, brain, and testis. The adenohypophysis maximally showed the ability to accumulate D-Asp when the latter is exogenously administered. In vivo experiments, consisting of the i.p. injection of D-Asp, showed that D-Asp induced both growth hormone and luteinizing hormone (LH) release. However, in vitro experiments showed that D-Asp was able to induce LH release from adenohypophysis only when this gland was co-incubated with the hypothalamus. This is because D-Asp also induces the release of GnRH from the hypothalamus, which in turn is directly responsible for the D-Asp-induced LH secretion from the pituitary gland. Compared to D-Asp, NMDA elicits its hormone release action at concentrations approximately 100-fold lower than D-Asp. D-AP5, a specific NMDA receptor antagonist, inhibited D-Asp and NMDA hormonal activity, demonstrating that these actions are mediated by NMDA receptors. NMDA is biosynthesized from D-Asp by an S-adenosylmethionine-dependent enzyme, which we tentatively denominated as NMDA synthase.

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Section: Methods 1: Enzymatic Fluorometricmentioning

confidence: 99%

Occurrence of D‐aspartic acid and N‐methyl‐D‐aspartic acid in rat neuroendocrine tissues and their role in the modulation of luteinizing hormone and growth hormone release

D‧Aniello¹,

et al. 2000

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“…[30] Instead, pronase from Streptomyces griseus was successfully used to cleave the peptide bond. [31,32] The resulting amino acid solution was then directly derivatized with ethyl chloroformate (ECF). [33,34] This derivatization can be performed in a slightly basic aqueous medium, thus avoiding any potentially difficult extraction steps prior to derivatization.…”

Section: Resultsmentioning

confidence: 99%

How to Spoil the Taste of Insect Prey? A Novel Feeding Deterrent against Ants Released by Larvae of the Alder Leaf Beetle, Agelastica alni

et al. 2010

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Chemical defense of leaf beetle larvae (Chrysomelidae) against enemies is provided by secretions containing a wide range of deterrent compounds or by unpalatable hemolymph constituents. Here we report a new, very strong feeding deterrent against ants released by larvae of the alder leaf beetle Agelastica alni when attacked. The larvae release a defensive fluid from openings of pairwise, dorsolaterally located tubercles on the first to the eighth abdominal segments. The fluid, consisting of hemolymph and probably a glandular cell secretion, has previously been shown to contain a very stable, non-volatile feeding deterrent. The major deterrent component was isolated by repeated HPLC separation and analyzed by NMR and MS. The compound proved to be gamma-L-glutamyl-L-2-furylalanine (1), a novel dipeptide containing the unusual amino acid L-2-furylalanine. This amino acid, although synthetically well known, has not previously been reported from natural sources. The absolute configuration of the natural compound was elucidated by enantioselective gas chromatography after derivatization. The structure of the dipeptide was verified by the synthesis of several isomeric dipeptides. In bioassays a concentration of 1 microg microL(-1) was sufficient to deter polyphagous Myrmica rubra ants from feeding.

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“…Partial chemical hydrolysis of proteins and peptides with hot 6 M HCl, followed by enzymatic hydrolysis with pronase, leucine aminopeptidase and peptidyl D-amino acid hydrolase, avoids racemization of the amino acids 281 . The problems arising from optical rotation measurements of chiral purity were reviewed.…”

Section: Chiral Puritymentioning

confidence: 99%

Analytical Aspects of Amino, Quaternary Ammonium, Nitro, Nitroso and Related Functional Groups

Zabicky

Bittner

2009

Patai's Chemistry of Functional Groups

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Improved Method for Hydrolyzing Proteins and Peptides Without Inducing Racemization and for Determining Their True D-Amino Acid Content

Cited by 28 publications

References 0 publications

Occurrence of D‐aspartic acid and N‐methyl‐D‐aspartic acid in rat neuroendocrine tissues and their role in the modulation of luteinizing hormone and growth hormone release

Occurrence of D‐aspartic acid and N‐methyl‐D‐aspartic acid in rat neuroendocrine tissues and their role in the modulation of luteinizing hormone and growth hormone release

How to Spoil the Taste of Insect Prey? A Novel Feeding Deterrent against Ants Released by Larvae of the Alder Leaf Beetle, Agelastica alni

Analytical Aspects of Amino, Quaternary Ammonium, Nitro, Nitroso and Related Functional Groups

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