Sequence analysis of phosphoserine‐containing peptides

Meyer, Helmut E.; Hoffmann-Posorske, Edeltraut; Korte, H.; Heilmeyer, Ludwig

doi:10.1016/0014-5793(86)81388-6

Cited by 262 publications

(178 citation statements)

References 13 publications

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“…The phosphate group of phosphothreonine is eliminated completely during the Edman degradation process, like the phosphate group of phosphoserine [24,25]. Therefore, the amount of DTT-addition products of the dehydroaminobutyric acid is increased compared with an unphosphorylated threonine.…”

Section: Discussionmentioning

confidence: 99%

N‐terminal sequence analysis of the 8 kDa protein in Chlamydomonas reinhardii Localization of the phosphothreonine

Dedner

Meyer

Ashton³

et al. 1988

FEBS Letters

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A phosphorylated 8 kDa protein of Chlamydomonas reinhurdii thylakoids has been isolated and its N-terminal amino acid sequence determined by gas-phase sequencing. The sequence analysis of the 48 amino acid residues revealed that this protein is about 50% homologous to the psb H gene products of higher plants. In contrast to them, it contains an insert of seven amino acid residues (Ser-5 to Lys-1 1). The first threonine residue was phosphorylated as determined by 32P detection during sequencing and also by analysis of the modified degradation products in the chemical reaction of the Edman degradation process. This latter method allows the identification of phosphorylated threonine residues without radiolabelling the protein. (Chlamydomonus reinhardiz)

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Section: Discussionmentioning

confidence: 99%

N‐terminal sequence analysis of the 8 kDa protein in Chlamydomonas reinhardii Localization of the phosphothreonine

Dedner

Meyer

Ashton³

et al. 1988

FEBS Letters

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“…Especially b-elimination of the phosphate moieties of Ser/Thr-phosphorylated proteins and peptides by alkali treatment and subsequent Michael-type addition of nucleophiles has a long standing record [34]. The first time this procedure was used for introduction of a biotin-containing tag with subsequent affinity purification of the peptides was reported by Oda et al in 2001 [35].…”

Section: Detection Of Phosphoproteins In 2-d Gelsmentioning

confidence: 99%

“…Since many phosphorylations are labile under Edman-conditions and sensitivity for the analysis of O-phosphates is rather poor, different applications involving b-elimination and subsequent Michael-type addition [34] or radiosequencing [75] have been developed.…”

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confidence: 99%

State-of-the-art in phosphoproteomics

2005

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Presently, phosphorylation of proteins is the most studied and best understood PTM. However, the analysis of phosphoproteins and phosphopeptides is still one of the most challenging tasks in contemporary proteome research. Since not every phosphoprotein is accessible by a certain method and identification of the phosphorylated amino acid residue is required in the majority of cases, various strategies for the detection and localization of phosphorylations have been developed. Identification and localization of protein phosphorylations is mostly done by MS nowadays but phosphoproteins and -peptides are often suppressed in comparison to the unphosphorylated species if measured in complex mixtures. Thus, the isolation of pure phosphopeptide samples is a main task. This review gives an overview over the most frequently used methods in isolation and detection of phosphoproteins and -peptides such as specific enrichment or separation strategies as well as the localization of the phosphorylated residues by various mass spectrometric techniques.

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“…According to the results presented in Since carbohydrate residues bound to serine only partially undergo f-elimination during Edman degradation, the relatively low yield of serine in positions 5 and 10 is most probably due to glycosylation. If phosphoserine residues were present in these positions, a quantitative yield of dithiothreitol-serine would have been observed (Meyer et al, 1986). In both potential glycosylation sites, glycine is present next to serine.…”

Section: Characterization Of the Pgmentioning

confidence: 99%

Purification and N-terminal amino acid sequence of a chondroitin sulphate/dermatan sulphate proteoglycan isolated from intima/media preparations of human aorta

Stöcker

Meyer

Wagener

et al. 1991

Biochemical Journal

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A proteoglycan (PG) was purified to homogeneity from intima/media preparations of human aorta specimens by the following chromatographic steps: Sepharose Q anion exchange, Sepharose CL-4B size exclusion, hydroxyapatite, MonoQ anion exchange and TSK G 4000 SW size exclusion. The purity of the preparation was established by SDS/PAGE using direct staining by silver or Dimethylmethylene Blue, as well as by Western blots of biotin-labelled samples. The electrophoretic mobility of the native PG was less than that of a 200000-Mr standard protein. After treatment with chondroitin sulphate lyase ABC, a core protein of Mr 15000 was revealed. The Mr of the glycosaminoglycan (GAG) peptides was less than 24000, by comparison with a keratan sulphate peptide. The composition of the GAG chains was determined by differential digestion of the PG by chondroitin sulphate lyases AC/ABC or chondroitin sulphate lyase AC alone followed by anion-exchange chromatography of the resulting disaccharides. The GAG chains are composed of approximately one-third of dermatan sulphate and two-thirds chondroitin sulphate disaccharide units. The sequence of the 20 N-terminal amino acids is identical with the sequence previously reported for PG I isolated from human developing bone [Fisher, Termine & Young (1989) J. Biol. Chem. 264, 4571-4576]. The assignment of glycosylation sites to the serine residues in positions 5 and 10 was confirmed. The findings indicate that the chondroitin sulphate/dermatan sulphate PG is a major PG in intima/media preparations of human aorta and represents a biglycan-type PG.

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Sequence analysis of phosphoserine‐containing peptides

Cited by 262 publications

References 13 publications

N‐terminal sequence analysis of the 8 kDa protein in Chlamydomonas reinhardii Localization of the phosphothreonine

N‐terminal sequence analysis of the 8 kDa protein in Chlamydomonas reinhardii Localization of the phosphothreonine

State-of-the-art in phosphoproteomics

Purification and N-terminal amino acid sequence of a chondroitin sulphate/dermatan sulphate proteoglycan isolated from intima/media preparations of human aorta

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