Topology of the Neurospora plasma membrane H(+)-ATPase. Localization of a transmembrane segment.
The Neurospora plasma membrane H(+)-ATPase is a polytopic integral membrane protein. To localize transmembrane segments, mutants were constructed that contained the amino and carboxyl termini of the H(+)-ATPase with putative transmembrane segment. A stretch of amino acid residues from yeast invertase that has three consensus N-linked glycosylation sites was placed carboxyl terminal of the putative transmembrane segment. RNA transcripts of these mutants were translated in a Neurospora in vitro system that was supplemented with microsomes from Neurospora. By the criteria of glycosylation of the polypeptide chain, resistance to extraction at pH 11.5, and protection from proteinase K digestion, only one transmembrane segment could be identified within the amino acid residues 272-314 of the primary sequence of the H(+)-ATPase.
Lin
J
J
Department of Biochemistry, University of Tennessee, Memphis 38163.
Addison
R
R
D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure; D003001:Cloning, Molecular; D016384:Consensus Sequence; D006031:Glycosylation; D008969:Molecular Sequence Data; D009491:Neurospora / Q000201:enzymology*; D014176:Protein Biosynthesis; D006180:Proton-Translocating ATPases / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D012331:RNA, Fungal / Q000096:biosynthesis / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D015183:Restriction Mapping; D014158:Transcription, Genetic
D012331:RNA, Fungal; D011993:Recombinant Fusion Proteins; D006180:Proton-Translocating ATPases