Synthesis of prothymosin alpha deduced from nucleotide sequence of the murine cDNA and its effect on the impaired T lymphocytes of uremic patients.
The complete murine prothymosin alpha molecule (110 residues) except for the N-terminal methionine deduced from the cloned cDNA has been synthesized by a solid-phase method. Peptide synthesis was performed manually by the stepwise solid-phase method using the base-labile Fmoc group for protecting the alpha-amino group. The peptide was assembled on a p-alkoxybenzyl alcohol resin. After the last coupling step, the Fmoc group was removed with 50% piperidine in DMF. The peptide resin was treated with thioanisole-o-cresol in TFA, and then purified by gel filtration, ion-exchange column chromatography and high-performance liquid chromatography. A 2.9-mg sample of a highly purified peptide was finally obtained. The overall yield of the synthesis was less than 1%, based on the amino acid content of the starting Fmoc-Asp (OtBu)-resin. The synthetic peptide was found to have a restoring activity on low-E-rosette-forming lymphocytes after incubation of peripheral blood from uremic patients with the synthetic peptide. This peptide exhibited far stronger restoring effect than that of our synthetic thymosin alpha 1.
Abiko
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T
Kidney Research Laboratory, Sendai, Japan.
Sekino
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H
D000595:Amino Acid Sequence; D000818:Animals; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000138:chemical synthesis / Q000302:isolation & purification / Q000494:pharmacology*; D012397:Rosette Formation; D013601:T-Lymphocytes / Q000276:immunology*; D013947:Thymosin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000302:isolation & purification / Q000494:pharmacology; D014511:Uremia / Q000276:immunology*
D018076:DNA, Complementary; D011498:Protein Precursors; C040782:prothymosin alpha; D013947:Thymosin