Liquid chromatographic analysis of phosphoamino acids at femtomole level using chemical derivatization with N-hydroxysuccinimidyl fluorescein-O-acetate

Deng, Yunsheng; Li, Rongjun; Zhang, Hua‐Shan; Du, Xiao-Lan; Wang, Hong

doi:10.1016/j.aca.2007.08.023

Cited by 9 publications

(4 citation statements)

References 34 publications

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“…Phosphoamino acids were derivatized with N ‐hydroxysuccinimidyl fluorescein‐ O ‐acetate (SIFA) at 40°C for 40 min in borate buffer (pH 8.5). SIFA fluoresces at 488 nm and LODs in the nanomolar range were observed 18. NDA (added with KCN), a well known primary amine derivatizing agent which is fluorogenic 19 was used for baseline separation of phosphothreonine, phosphotyrosine and phosphoserine (LODs 7.0, 5.6 and 7.2 nM, respectively) in 180 s and validated with β‐casein hydrolysate 20.…”

Section: New Ce Methodsmentioning

confidence: 99%

Recent advances in amino acid analysis by CE

Poinsot

Gavard

Feurer³

et al. 2009

Electrophoresis

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This article describes the most important articles that have been published on amino acid analysis using CE during the period from June 2007 to May 2009. It follows the format of the previous articles of Smith [Electrophoresis 1999, 20, 3078-3083], Prata et al. [Electrophoresis 2001, 22, 4129-4138] and Poinsot et al. [Electrophoresis 2003, 24, 4047-4062; Electrophoresis 2006, 27, 176-194; Electrophoresis 2008, 29, 207-223]. For several years we have presented the new developments in amino acid analysis with CE which describe the use of laser emitting diodes for LIF as well as via MS. In addition, we describe articles concerning clinical studies and neuroclinical applications.

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Section: New Ce Methodsmentioning

confidence: 99%

Recent advances in amino acid analysis by CE

Poinsot

Gavard

Feurer³

et al. 2009

Electrophoresis

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“…These approaches increase the abundance of phosphorylated peptides or proteins in LC-MS/MS assays. These enrichment methods include pSer/pThr/pTyr phosphorylation-specific antibody-based affinity capture ( Stoevesandt and Taussig, 2013 ), chemical derivatization of phosphorylated AAs ( Deng et al, 2007 ), metal ion-based affinity capture ( Blacken et al, 2008 ), ion-exchange chromatography ( Jungbauer and Hahn, 2009 ), and classical immunoprecipitation technology using phosphorylation-specific antibodies ( Kurosaki et al, 2019 ). The enriched proteins are hydrolyzed to form peptides via trypsin digestion in vitro , and LC-MS/MS analyses are performed to detect the phosphorylated sites.…”

Section: Common Assays For Identification Of Phosphorylated Plant Vir...mentioning

confidence: 99%

Phosphorylation of plant virus proteins: Analysis methods and biological functions

Zhuang

Guo²,

Gu³

et al. 2022

Front. Microbiol.

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Phosphorylation is one of the most extensively investigated post-translational modifications that orchestrate a variety of cellular signal transduction processes. The phosphorylation of virus-encoded proteins plays an important regulatory role in the infection cycle of such viruses in plants. In recent years, molecular mechanisms underlying the phosphorylation of plant viral proteins have been widely studied. Based on recent publications, our study summarizes the phosphorylation analyses of plant viral proteins and categorizes their effects on biological functions according to the viral life cycle. This review provides a theoretical basis for elucidating the molecular mechanisms of viral infection. Furthermore, it deepens our understanding of the biological functions of phosphorylation in the interactions between plants and viruses.

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“…A number of analytical techniques have been exploited for phosphoamino acids analysis, as found in two review articles 4, 5, such as radioactive labeling 6, antibody recognition 7, HPLC 3, 8–10, GC 11, 12, CE 13–20, and MS 21. In comparison to other methods, CE represents a sensitive and high‐resolution separation method that is well suited for the analysis of trace levels of phosphoamino acids in biological samples due to its rapid analysis times, minimal band dispersion, and small sample requirements.…”

Section: Introductionmentioning

confidence: 99%

Rapid analysis of phosphoamino acids from phosvitin by near‐infrared cyanine 1‐(ε‐succinimydyl‐hexanoate)‐1′‐methyl‐3,3,3′,3′‐tetramethyl‐indocarbocyanine‐5,5′‐disulfonate potassium derivatization and polyacrylamide‐coated CE with LIF detection

Wang

Zhang

2011

Electrophoresis

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A polyacrylamide-coated CE method with LIF detection was developed for analyzing three phosphoamino acids including phosphotyrosine (p-Tyr), phosphothreonine (p-Thr), and phosphoserine (p-Ser). A near-infrared dye, 1-(ε-succinimydyl-hexanoate)-1'-methyl-3,3,3',3'-tetramethyl-indocarbocyanine-5,5'-disulfonate potassium (MeCy5-OSu) was employed for derivatization of these phosphoamino acids. Results indicated that the complete baseline resolution of each phosphoamino acid was obtained within 6.1 min, using 10 mmol/L phosphate buffer (pH 4.0) containing 60 mmol/L SDS as running buffer. The highest derivatization efficiency was achieved in 0.2 mol/L borate buffer (pH 8.8) for 30 min at 30 °C. Linearity of response was found in the range of 0.05-1 μmol/L. The correlation coefficients for these phosphoamino acids were from 0.9940 to 0.9976. The LODs for phosphotyrosine, phosphothreonine, and phosphoserine were about 6, 8, and 8 nmol/L, respectively. The proposed method has been successfully applied to the determination of phosphoamino acids in the hydrolysis sample of a phosphorylated phosvitin. Average recoveries for phosvitin sample were in the range of 94.0-98.0% and coefficients of variation ranged from 2.7 to 4.8%.

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Liquid chromatographic analysis of phosphoamino acids at femtomole level using chemical derivatization with N-hydroxysuccinimidyl fluorescein-O-acetate

Cited by 9 publications

References 34 publications

Recent advances in amino acid analysis by CE

Recent advances in amino acid analysis by CE

Phosphorylation of plant virus proteins: Analysis methods and biological functions

Rapid analysis of phosphoamino acids from phosvitin by near‐infrared cyanine 1‐(ε‐succinimydyl‐hexanoate)‐1′‐methyl‐3,3,3′,3′‐tetramethyl‐indocarbocyanine‐5,5′‐disulfonate potassium derivatization and polyacrylamide‐coated CE with LIF detection

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