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Biochemical fractionation and assembly of the membrane components that mediate nascent chain targeting and translocation.

Publication ,  Journal Article
Nicchitta, CV; Migliaccio, G; Blobel, G
Published in: Cell
May 17, 1991

Fractionation of a microsomal detergent extract with ammonium sulfate allows separation of the signal recognition particle receptor (SR alpha), which is required for targeting of the nascent chain, from other microsomal proteins, such as signal peptidase, whose activity is displayed during subsequent translocation. The reconstituted SR alpha-enriched fraction is functional in assays of precursor targeting and elongation arrest release but lacks translocation activity. This defect can be complemented by addition, prior to reconstitution, of a separate protein subfraction. In addition, protein components necessary for translocation can be reversibly depleted from the complementary fraction, under conditions where precursor targeting is retained, by sulfhydryl-directed chromatography. Thus, precursor binding and translocation can be biochemically uncoupled, indicating that they are sequential reactions mediated by distinct components.

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Published In

Cell

DOI

ISSN

0092-8674

Publication Date

May 17, 1991

Volume

65

Issue

4

Start / End Page

587 / 598

Location

United States

Related Subject Headings

  • Triticum
  • Signal Recognition Particle
  • Ribosomes
  • Ribonucleoproteins
  • Reticulocytes
  • Receptors, Peptide
  • Receptors, Cytoplasmic and Nuclear
  • Receptors, Cell Surface
  • RNA, Messenger
  • Protein Precursors
 

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Nicchitta, C. V., Migliaccio, G., & Blobel, G. (1991). Biochemical fractionation and assembly of the membrane components that mediate nascent chain targeting and translocation. Cell, 65(4), 587–598. https://doi.org/10.1016/0092-8674(91)90091-c
Nicchitta, C. V., G. Migliaccio, and G. Blobel. “Biochemical fractionation and assembly of the membrane components that mediate nascent chain targeting and translocation.Cell 65, no. 4 (May 17, 1991): 587–98. https://doi.org/10.1016/0092-8674(91)90091-c.
Nicchitta, C. V., et al. “Biochemical fractionation and assembly of the membrane components that mediate nascent chain targeting and translocation.Cell, vol. 65, no. 4, May 1991, pp. 587–98. Pubmed, doi:10.1016/0092-8674(91)90091-c.
Journal cover image

Published In

Cell

DOI

ISSN

0092-8674

Publication Date

May 17, 1991

Volume

65

Issue

4

Start / End Page

587 / 598

Location

United States

Related Subject Headings

  • Triticum
  • Signal Recognition Particle
  • Ribosomes
  • Ribonucleoproteins
  • Reticulocytes
  • Receptors, Peptide
  • Receptors, Cytoplasmic and Nuclear
  • Receptors, Cell Surface
  • RNA, Messenger
  • Protein Precursors