Segregation of mutant ovalbumins and ovalbumin-globin fusion proteins in Xenopus oocytes. Identification of an ovalbumin signal sequence

L. Tabe, Paul A Krieg, R. Strachan, D. Jackson, E. Wallis, A. Colman

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

The intramolecular signals for chicken ovalbumin secretion were examined by producing mutant proteins in Xenopus oocytes. An ovalbumin complementary DNA clone was manipulated in vitro, and constructs containing altered protein-coding sequences and either the simian virus 40 (SV40) early promoter or Herpes simplex thymidine kinase promoter, were microinjected into Xenopus laevis oocytes. The removal of the eight extreme N-terminal amino acids of ovalbumin had no effect on the segregation of ovalbumin with oocyte membranes nor on its secretion. A protein lacking amino acids 2 to 21 was sequestered in the endoplasmic reticulum but remained strongly associated with the oocyte membranes rather than being secreted. Removal of amino acids 231 to 279, a region previously reported to have membrane-insertion function, resulted in a protein that also entered the endoplasmic reticulum but was not secreted. Hybrid proteins containing at their N terminus amino acids 9 to 41 or 22 to 41 of ovalbumin fused to the complete chimpanzee α-globin polypeptide were also sequestered by oocyte membranes. We conclude that the ovalbumin "signal" sequeǹce is internally located within amino acids 22 to 41, and we speculate that amino acids 9 to 21 could be important for the completion of ovalbumin translocation through membranes.

Original languageEnglish (US)
Pages (from-to)645-666
Number of pages22
JournalJournal of Molecular Biology
Volume180
Issue number3
DOIs
StatePublished - Dec 15 1984
Externally publishedYes

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Xenopus Proteins
Globins
Ovalbumin
Protein Sorting Signals
Oocytes
Amino Acids
Membranes
Endoplasmic Reticulum
Proteins
Herpes Simplex
Simian virus 40
Pan troglodytes
Thymidine Kinase
Xenopus laevis
Mutant Proteins
Xenopus
Chickens
Complementary DNA
Clone Cells
Peptides

ASJC Scopus subject areas

  • Virology

Cite this

Segregation of mutant ovalbumins and ovalbumin-globin fusion proteins in Xenopus oocytes. Identification of an ovalbumin signal sequence. / Tabe, L.; Krieg, Paul A; Strachan, R.; Jackson, D.; Wallis, E.; Colman, A.

In: Journal of Molecular Biology, Vol. 180, No. 3, 15.12.1984, p. 645-666.

Research output: Contribution to journalArticle

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