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  • Acid hydrolysis of silk fibroins and determination of the enrichment of isotopically labeled amino acids using precolumn derivatization and high-performance liquid chromatography-electrospray ionization-mass spectrometry.

Acid hydrolysis of silk fibroins and determination of the enrichment of isotopically labeled amino acids using precolumn derivatization and high-performance liquid chromatography-electrospray ionization-mass spectrometry.

Analytical biochemistry (2002-11-21)
Sonja Hess, Jacco van Beek, Lewis K Pannell
摘要

Silk fibroins from moth larvae and spiders are composed of highly repetitive Ala- and Gly-rich blocks that determine their structure, properties, and function. To investigate the metabolic integration of isotopically labeled amino acids in the excreted silk, the enrichment of ingested tracers was determined after acid hydrolysis of the fibroins. Thus, spiders and moth larvae were fed with stable isotope tracers such as [1-13C]Ala or [1-13C]Gly and silked. After hydrolysis of the silk proteins, the corresponding amino acids were derivatized with Nalpha-(2,4-dinitro-5-fluorophenyl)-L-alaninamide (Marfey's reagent) and separated by liquid chromatography. The isotopical enrichment of the amino acids was determined by online electrospray mass spectrometry and calculated by newly developed software. Depending on the feeding protocol, enrichments of up to 58% in Gly and 31% in Ala were found in the investigated silks. The highly enriched silk fibroins are suitable for further structural investigation such as solid-state nuclear magnetic resonance.

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Sigma-Aldrich
Nα-(2,4-二硝基-5-氟苯基)-L氨基丙酰胺, powder
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2 g
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100 g
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500 g
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Supelco
Nα-(2,4-二硝基-5-氟苯基)-L氨基丙酰胺, derivatization grade (chiral), LiChropur, ≥99.0%
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货号包装规格是否有货价格数量
2 g
有货
详情...
¥325.76
100 g
有货
详情...
¥448.69
500 g
有货
详情...
¥1,207.47
2 kg
预计发货时间 2025年8月14日
详情...
¥4,107.83