Import of Honeybee Prepromelittin into the Endoplasmic Reticulum: Structural Basis for Independence of SRP and Docking Protein

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1987 Jul 1

PMID 2820722

Citations 42

Authors

G Muller

R Zimmermann

Affiliations

Soon will be listed here.

Abstract

Honeybee prepromelittin is correctly processed and imported by dog pancreas microsomes. Insertion of prepromelittin into microsomal membranes, as assayed by signal sequence removal, does not depend on signal recognition particle (SRP) and docking protein. We addressed the question as to how prepromelittin bypasses the SRP/docking protein system. Hybrid proteins between prepromelittin, or carboxy-terminally truncated derivatives, and the cytoplasmic protein dihydrofolate reductase from mouse were constructed. These hybrid proteins were analysed for membrane insertion and sequestration into microsomes. The results suggest the following: (i) The signal sequence of prepromelittin is capable of interacting with the SRP/docking protein system, but this interaction is not mandatory for membrane insertion; this is related to the small size of prepromelittin. (ii) In prepromelittin a cluster of negatively charged amino acids must be balanced by a cluster of positively charged amino acids in order to allow membrane insertion. (iii) In general, a signal sequence can be sufficient to mediate membrane insertion independently of SRP and docking protein in the case of short precursor proteins; however, the presence and distribution of charged amino acids within the mature part of these precursors can play distinct roles.

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References

Zimmermann R, Mollay C . Import of honeybee prepromelittin into the endoplasmic reticulum. Requirements for membrane insertion, processing, and sequestration. J Biol Chem. 1986; 261(27):12889-95. View

Kuhn A, Wickner W, KREIL G . The cytoplasmic carboxy terminus of M13 procoat is required for the membrane insertion of its central domain. Nature. 1986; 322(6077):335-9. DOI: 10.1038/322335a0. View

Kuhn A, KREIL G, Wickner W . Both hydrophobic domains of M13 procoat are required to initiate membrane insertion. EMBO J. 1986; 5(13):3681-5. PMC: 1167410. DOI: 10.1002/j.1460-2075.1986.tb04699.x. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Radloff R, BAUER W, Vinograd J . A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proc Natl Acad Sci U S A. 1967; 57(5):1514-21. PMC: 224502. DOI: 10.1073/pnas.57.5.1514. View

Schlenstedt G, Zimmermann R . Import of frog prepropeptide GLa into microsomes requires ATP but does not involve docking protein or ribosomes. EMBO J. 1987; 6(3):699-703. PMC: 553453. DOI: 10.1002/j.1460-2075.1987.tb04810.x. View

Blobel G, Sabatini D . Controlled proteolysis of nascent polypeptides in rat liver cell fractions. I. Location of the polypeptides within ribosomes. J Cell Biol. 1970; 45(1):130-45. PMC: 2107986. DOI: 10.1083/jcb.45.1.130. View

Mandel M, Higa A . Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970; 53(1):159-62. DOI: 10.1016/0022-2836(70)90051-3. View

Kedes L, Chang A, Houseman D, Cohen S . Isolation of histone genes from unfractionated sea urchin DNA by subculture cloning in E. coli. Nature. 1975; 255(5509):533-8. DOI: 10.1038/255533a0. View

10.

Blobel G, Dobberstein B . Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975; 67(3):835-51. PMC: 2111658. DOI: 10.1083/jcb.67.3.835. View

11.

Pelham H, Jackson R . An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976; 67(1):247-56. DOI: 10.1111/j.1432-1033.1976.tb10656.x. View

12.

McDonell M, Simon M, Studier F . Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977; 110(1):119-46. DOI: 10.1016/s0022-2836(77)80102-2. View

13.

SANGER F, Nicklen S, Coulson A . DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977; 74(12):5463-7. PMC: 431765. DOI: 10.1073/pnas.74.12.5463. View

14.

Jackson R, Blobel G . Post-translational cleavage of presecretory proteins with an extract of rough microsomes from dog pancreas containing signal peptidase activity. Proc Natl Acad Sci U S A. 1977; 74(12):5598-602. PMC: 431824. DOI: 10.1073/pnas.74.12.5598. View

15.

Suchanek G, KREIL G, Hermodson M . Amino acid sequence of honeybee prepromelittin synthesized in vitro. Proc Natl Acad Sci U S A. 1978; 75(2):701-4. PMC: 411324. DOI: 10.1073/pnas.75.2.701. View

16.

Zimmerman R, Paluch U, Sprinzl M, Neupert W . Cell-free synthesis of the mitochondrial ADP/ATP carrier protein of Neurospora crassa. Eur J Biochem. 1979; 99(2):247-52. DOI: 10.1111/j.1432-1033.1979.tb13251.x. View

17.

Birnboim H, DOLY J . A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979; 7(6):1513-23. PMC: 342324. DOI: 10.1093/nar/7.6.1513. View

18.

Ito K, Date T, Wickner W . Synthesis, assembly into the cytoplasmic membrane, and proteolytic processing of the precursor of coliphage M13 coat protein. J Biol Chem. 1980; 255(5):2123-30. View

19.

Nunberg J, Kaufman R, Chang A, Cohen S, Schimke R . Structure and genomic organization of the mouse dihydrofolate reductase gene. Cell. 1980; 19(2):355-64. DOI: 10.1016/0092-8674(80)90510-3. View

20.

Chamberlain J . Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate. Anal Biochem. 1979; 98(1):132-5. DOI: 10.1016/0003-2697(79)90716-4. View