Targeting and topology in the membrane of plant 3-hydroxy-3-methylglutaryl coenzyme A reductase.
The enzyme 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) catalyzes the synthesis of mevalonate. This is the first committed step of isoprenoid biosynthesis. A common feature of all known plant HMGR isoforms is the presence of two highly conserved hydrophobic sequences in the N-terminal quarter of the protein. Using an in vitro system, we showed that the two hydrophobic sequences of Arabidopsis HMGR1S function as internal signal sequences. Specific recognition of these sequences by the signal recognition particle mediates the targeting of the protein to microsomes derived from the endoplasmic reticulum. Arabidopsis HMGR is inserted into the microsomal membrane, and the two hydrophobic sequences become membrane-spanning segments. The N-terminal end and the C-terminal catalytic domain of Arabidopsis HMGR are positioned on the cytosolic side of the membrane, whereas only a short hydrophilic sequence is exposed to the lumen. Our results suggest that the plant HMGR isoforms known to date are primarily targeted to the endoplasmic reticulum and have the same topology in the membrane. This reinforces the hypothesis that mevalonate is synthesized only in the cytosol. The possibility that plant HMGRs might be located in different regions of the endomembrane system is discussed.
Campos
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Departament de Bioquímica i Biologia Molecular, Universitat de Barcelona, Spain.
Boronat
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D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D017124:Conserved Sequence; D017931:DNA Primers; D018076:DNA, Complementary; D004721:Endoplasmic Reticulum / Q000201:enzymology; D006903:Hydroxymethylglutaryl CoA Reductases / Q000096:biosynthesis* / Q000737:chemistry*; D007425:Intracellular Membranes / Q000201:enzymology* / Q000648:ultrastructure; D007527:Isoenzymes / Q000096:biosynthesis / Q000737:chemistry; D008861:Microsomes / Q000201:enzymology*; D008961:Models, Structural; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D021382:Protein Sorting Signals / Q000378:metabolism; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D017384:Sequence Deletion; D014158:Transcription, Genetic
D017931:DNA Primers; D018076:DNA, Complementary; D007527:Isoenzymes; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D011994:Recombinant Proteins; D006903:Hydroxymethylglutaryl CoA Reductases