Fluorescent peptide mapping with microgram quantities of protein

Benson, James R.

doi:10.1016/s0003-2697(76)80012-7

Cited by 33 publications

(5 citation statements)

References 5 publications

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“…A control vial containing 500 pi. of water was treated with trypsin in exactly the same fashion as vials containing protein samples to determine if the sample handling procedure (Benson, 1976) prevented contamination of OPAreactive substances. Analysis of this control provided a background against which all peptide maps could be compared (Figure 1).…”

Section: Resultsmentioning

confidence: 99%

“…The Microbore Chromatograph. The microbore ion-exchange liquid chromatograph utilized constant-pressure eluent pumps modeled after those originally described by Hare (1969Hare ( , 1972 and is described in detail elsewhere (Benson, 1976). A 57 °C jacketed column containing a 0.20 X 25 cm bed of Durrum DC-4A cation-exchange resin was used (Durrum Chemical Corp., Sunnyvale, Calif.); eluent flow rate was 6.3 mL/h (linear flow velocity = 3.3 cm/min).…”

Section: Methodsmentioning

confidence: 99%

“…Sample Preparation. Preparation of purified proteins and the tryptic hydrolysis conditions are given in detail elsewhere (Benson, 1976). Typically, 100 pg of purified denatured protein diluted to 500 pL with water maintained at 37 °C was treated with a total of 4 pg of Tos-PheCHjCl trypsin (Worthington Biochemicals) at pH 8 to 9.…”

Section: Methodsmentioning

confidence: 99%

“…Starting with only microgram amounts of purified material, we found evidence of regions of relatedness among proteins reported to contain interspecies antigenic determinants and also in the 10 000 molecular weight proteins from feline and murine leukemia viruses, although these are reported to contain no common antigens (Green et al, 1973;Schafer et al, 1975). The technique, first described by Benson (1976), combines microbore ion-exchange chromatography with a new, highly sensitive fluorescent assay for biogenic amines. The fluorescent assay employs o-phthalaldehyde (OPA) which in the presence of 2-mercaptoethanol reacts with primary amines to form highly fluorescent products.…”

mentioning

confidence: 99%

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Interspecies homology of RNA tumor virus proteins

Benson¹,

Hayflick²

1977

Biochemistry

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We report the application of a highly sensitive column chromatographic technique to the comparison of tryptic peptide maps of some RNA tumor virus proteins. By combining microbore ion-exchange chromatography with a sensitive fluorescent assay using o-phthalaldehyde, we obtained high-resolution peptide maps starting with only microgram amounts of protein. Our discovery of coincident peptides from the 15,000 and 30,000 molecular weight proteins from murine and feline leukemia viruses supports serological evidence for interspecies antigenic determinants; coincident peptides were also found for the 10,000 molecular weight proteins from these viruses, although immunochemical data did not reveal interspecies determinants. The relatively large number of coeluting peptides found in the 15,000 and 10,000 molecular weight proteins is strong evidence for the existence of homology.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Interspecies homology of RNA tumor virus proteins

Benson¹,

Hayflick²

1977

Biochemistry

Self Cite

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“…Amino acid analvsis. Peptides were hydrolysed for 22h at 1 10°C in sealed tubes in vacuo, and hydrolysates were run on a Durrum D-500 amino acid analyser (Benson, 1976).…”

Section: Methodsmentioning

confidence: 99%

The primary structure of the constant region of Basilea-rabbit immunoglobulin λ-chains

García¹,

Jaton²

1981

Biochemical Journal

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The amino acid sequence of the constant region of the immunoglobulin lambda-chain of the rabbit (C lambda) has been determined. This chain was obtained from a homogeneous anti-(type II pneumococcal polysaccharide) antibody fraction raised in Basilea rabbit 4548. The C lambda-region sequence was established by the analysis of tryptic and some overlapping chymotryptic peptides and partly by homology with known C lambda-region sequences of lambda-chains from other species. Peptides were isolated by a combination of methods including high-voltage paper electrophoresis, gel filtration and high-pressure liquid chromatography. The peptides were subjected to automated Edman degradation in the presence of Polybrene. The sequence showed no evidence of heterogeneity. Large interspecies similarities, as well as amino acid interchanges, are apparent among various C lambda regions; the degrees of homology between rabbit C lambda region and the C lambda region from human, porcine and murine chains are 72.6, 71 and 72% respectively. The similarity of lambda-chains of different species is much greater than that found between rabbit kappa- and lambda-chains (about 34% homology). Genetic implications of these results are discussed.

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Peptide Mapping of Proteins

James

1980

Methods of Biochemical Analysis

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Fluorescent peptide mapping with microgram quantities of protein

Cited by 33 publications

References 5 publications

Interspecies homology of RNA tumor virus proteins

Interspecies homology of RNA tumor virus proteins

The primary structure of the constant region of Basilea-rabbit immunoglobulin λ-chains

Peptide Mapping of Proteins

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