Import of honeybee prepromelittin into the endoplasmic reticulum: structural basis for independence of SRP and docking protein.
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.
Müller
G
G
Institut für Physiologische Chemie der Universität München, FRG.
Zimmermann
R
R
D000818:Animals; D001514:Bee Venoms / Q000235:genetics*; D001516:Bees; D004247:DNA / Q000378:metabolism; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D008555:Melitten / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism*; D010179:Pancreas / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000235:genetics* / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism*; D018271:Signal Recognition Particle; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014158:Transcription, Genetic
D001514:Bee Venoms; D008565:Membrane Proteins; D011498:Protein Precursors; D012261:Ribonucleoproteins; D018271:Signal Recognition Particle; C503637:Srp72 protein, mouse; C488951:Srpr protein, mouse; D008555:Melitten; C016694:prepromelittin; D004247:DNA; D013762:Tetrahydrofolate Dehydrogenase