Export of honeybee prepromelittin in Escherichia coli depends on the membrane potential but does not depend on proteins secA and secY.
Honeybee prepromelittin (70 amino acid residues), the precursor of an eukaryotic secretory protein, and a hybrid protein between prepromelittin and mouse dihydrofolate reductase (257 amino acid residues) were expressed in Escherichia coli and characterized with respect to their requirements for transport across the plasma membrane. Both precursor proteins are posttranslationally processed and exported into the periplasm, and they both depend on the membrane potential for this to occur. With respect to dependence on components of the export machinery, however, the two precursor proteins show striking differences: the small precursor protein prepromelittin does not require the function of proteins secA and secY; the large precursor protein prepromelittin-dihydrofolate reductase, on the other hand, depends on both components. The implications of these observations with respect to the mechanisms of protein export in E. coli and of protein import into the endoplasmic reticulum are discussed.
Cobet
W W
WW
Institut für Physiologische Chemie der Universität München, Federal Republic of Germany.
Mollay
C
C
Müller
G
G
Zimmermann
R
R
D000818:Animals; D001426:Bacterial Proteins / Q000502:physiology*; D001514:Bee Venoms / Q000235:genetics*; D001516:Bees / Q000235:genetics*; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002462:Cell Membrane / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics* / Q000502:physiology; D007700:Kinetics; D008555:Melitten / Q000096:biosynthesis / Q000235:genetics*; D008564:Membrane Potentials; D010957:Plasmids; D011498:Protein Precursors / Q000096:biosynthesis / Q000235:genetics*; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics
D001426:Bacterial Proteins; D001514:Bee Venoms; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D008555:Melitten; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; C016694:prepromelittin; D013762:Tetrahydrofolate Dehydrogenase