Abstract
The eukaryotic translation initiation factor eIF-4D is the only protein known to contain the unusual amino acid hypusine, a posttranslationally modified lysine. For the production of monoclonal antibodies the hypusine-containing protein (HP) was isolated from Dictyostelium discoideum. Using these monoclonal antibodies, a full-length cDNA clone was isolated from a lambda gt11 library. The D. discoideum HP consists of 169 amino acids and has a molecular mass of 18.3 kDa. It is encoded by a single gene. Tryptic and cyanogen bromide peptides were prepared from the purified protein and sequenced. The hypusine residue is located at amino acid position 65 of the HP. The corresponding mRNA of approx. 0.6 kb is present throughout the life cycle of D. discoideum.
MeSH terms
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal
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Base Sequence
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Chromatography, Affinity
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Codon
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Cyanogen Bromide
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DNA / genetics*
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DNA Restriction Enzymes
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Dictyostelium / analysis*
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Dictyostelium / growth & development
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Dictyostelium / metabolism
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Electrophoresis, Polyacrylamide Gel
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Eukaryotic Translation Initiation Factor 5A
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Immunoassay
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Lysine / analogs & derivatives*
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Lysine / analysis
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Mice
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Mice, Inbred BALB C
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Molecular Sequence Data
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Molecular Weight
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Nucleic Acid Hybridization
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Peptide Fragments / analysis
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Peptide Initiation Factors* / analysis
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Peptide Initiation Factors* / genetics
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RNA, Messenger / metabolism
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RNA-Binding Proteins*
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Trypsin
Substances
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Antibodies, Monoclonal
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Codon
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Peptide Fragments
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Peptide Initiation Factors
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RNA, Messenger
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RNA-Binding Proteins
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hypusine
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DNA
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DNA Restriction Enzymes
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Trypsin
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Lysine
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Cyanogen Bromide