Localization of the intrachain disulfide bonds of the envelope glycoprotein 71 from Friend murine leukemia virus.
Envelope glycoprotein 71 from Friend murine leukemia virus was purified to homogeneity by reversed-phase HPLC. It could be shown that all 20 cysteine residues of the molecule are linked by disulfide bonds. After complete tryptic digestion, peptides containing cystine were identified by comparison of the reversed-phase HPLC profile of the digest with that of a reduced aliquot which had been subjected to affinity chromatography on thiol-Sepharose. The locations of the 10 disulfide bonds were determined by isolation, further digestion and analysis of peptides containing cystine. The first cysteine residue of the sequence (Cys46) was shown to be coupled to the sixth (Cys98), leading to a large loop containing four additional cysteine residues. Computer model building and energy calculations led to the assignment of Cys72 to Cys87 and Cys73 to Cys83. The following four cysteine residues of the sequence also constitute a structural unit, with Cys121 bonded to Cys141 and Cys133 to Cys146, and the last two cysteine residues in the amino-terminal domain of glycoprotein 71 form a small loop (Cys178 to Cys184). The first two cysteine residues of the carboxy-terminal domain produce a very small hydrophobic loop (Cys312-Cys315). Cys361 is bound to Cys373, Cys342 to Cys396 and Cys403 to Cys416. A model for the folding pattern of the viral glycoprotein is proposed.
Linder
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Biochemisches Institut am Klinikum, Justus-Liebig-Universität, Giessen, Federal Republic of Germany.
Linder
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Hahnen
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J
Schott
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HH
Stirm
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D000595:Amino Acid Sequence; D002846:Chromatography, Affinity; D003553:Cystine / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005622:Friend murine leukemia virus / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011487:Protein Conformation; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics / Q000378:metabolism*; D014357:Trypsin / Q000737:chemistry; D014759:Viral Envelope Proteins
D004220:Disulfides; D015682:Retroviridae Proteins, Oncogenic; D014759:Viral Envelope Proteins; C038947:glycoprotein 71, Friend murine leukemia virus; D003553:Cystine; D014357:Trypsin