Determinants of Membrane Protein Topology

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1987 Dec 1

PMID 3317413

Citations 131

Authors

D Boyd

C Manoil

J Beckwith

Affiliations

Soon will be listed here.

Abstract

The topology of the integral membrane protein MalF, which is required for maltose transport in Escherichia coli, has been analyzed using fusions of alkaline phosphatase (EC 3.1.3.1). The properties of such fusion strains support a MalF structure previously proposed on theoretical grounds. Several transmembrane segments within MalF can act as signal sequences in exporting alkaline phosphatase. Other transmembrane sequences, in conjunction with cytoplasmic domains, can stably anchor alkaline phosphatase in the cytoplasm. Our results suggest that features of the amino acid sequence (possibly the positively charged amino acids) of the cytoplasmic domains of membrane proteins are important in anchoring these domains in the cytoplasm. These studies in conjunction with our earlier results show that alkaline phosphatase fusions to membrane proteins can be an important aid in analyzing membrane topology and its determinants.

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References

MALAMY M, HORECKER B . RELEASE OF ALKALINE PHOSPHATASE FROM CELLS OF ESCHERICHIA COLI UPON LYSOZYME SPHEROPLAST FORMATION. Biochemistry. 1964; 3:1889-93. DOI: 10.1021/bi00900a017. View

Roberts C, Chlebowski J . Trypsin modification of Escherichia coli alkaline phosphatase. J Biol Chem. 1984; 259(2):729-33. View

Zagursky R, Berman M . Cloning vectors that yield high levels of single-stranded DNA for rapid DNA sequencing. Gene. 1984; 27(2):183-91. DOI: 10.1016/0378-1119(84)90139-2. View

Froshauer S, Beckwith J . The nucleotide sequence of the gene for malF protein, an inner membrane component of the maltose transport system of Escherichia coli. Repeated DNA sequences are found in the malE-malF intercistronic region. J Biol Chem. 1984; 259(17):10896-903. View

Heijne G . The distribution of positively charged residues in bacterial inner membrane proteins correlates with the trans-membrane topology. EMBO J. 1986; 5(11):3021-7. PMC: 1167256. DOI: 10.1002/j.1460-2075.1986.tb04601.x. View

Geider K, Hohmeyer C, Haas R, Meyer T . A plasmid cloning system utilizing replication and packaging functions of the filamentous bacteriophage fd. Gene. 1985; 33(3):341-9. DOI: 10.1016/0378-1119(85)90242-2. View

Gutierrez C, BARONDESS J, Manoil C, Beckwith J . The use of transposon TnphoA to detect genes for cell envelope proteins subject to a common regulatory stimulus. Analysis of osmotically regulated genes in Escherichia coli. J Mol Biol. 1987; 195(2):289-97. DOI: 10.1016/0022-2836(87)90650-4. View

Enea V, Zinder N . Interference resistant mutants of phage f1. Virology. 1982; 122(1):222-6. DOI: 10.1016/0042-6822(82)90395-6. View

Michaelis S, Inouye H, Oliver D, Beckwith J . Mutations that alter the signal sequence of alkaline phosphatase in Escherichia coli. J Bacteriol. 1983; 154(1):366-74. PMC: 217468. DOI: 10.1128/jb.154.1.366-374.1983. View

10.

Mueckler M, Lodish H . The human glucose transporter can insert posttranslationally into microsomes. Cell. 1986; 44(4):629-37. DOI: 10.1016/0092-8674(86)90272-2. View

11.

Silhavy T, Brickman E, Bassford Jr P, Casadaban M, Shuman H, Schwartz V . Structure of the malB region in Escherichia coli K12. II. Genetic map of the malE,F,G operon. Mol Gen Genet. 1979; 174(3):249-59. DOI: 10.1007/BF00267797. View

12.

Brosius J, Holy A . Regulation of ribosomal RNA promoters with a synthetic lac operator. Proc Natl Acad Sci U S A. 1984; 81(22):6929-33. PMC: 392049. DOI: 10.1073/pnas.81.22.6929. View

13.

Manoil C, Beckwith J . A genetic approach to analyzing membrane protein topology. Science. 1986; 233(4771):1403-8. DOI: 10.1126/science.3529391. View

14.

Craik C, Largman C, Fletcher T, Roczniak S, Barr P, Fletterick R . Redesigning trypsin: alteration of substrate specificity. Science. 1985; 228(4697):291-7. DOI: 10.1126/science.3838593. View

15.

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

16.

Johnson R, Reznikoff W . Copy number control of Tn5 transposition. Genetics. 1984; 107(1):9-18. PMC: 1202317. DOI: 10.1093/genetics/107.1.9. View

17.

Chang A, Cohen S . Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978; 134(3):1141-56. PMC: 222365. DOI: 10.1128/jb.134.3.1141-1156.1978. View

18.

Wolfe P, Rice M, Wickner W . Effects of two sec genes on protein assembly into the plasma membrane of Escherichia coli. J Biol Chem. 1985; 260(3):1836-41. View

19.

Michaelis S, HUNT J, Beckwith J . Effects of signal sequence mutations on the kinetics of alkaline phosphatase export to the periplasm in Escherichia coli. J Bacteriol. 1986; 167(1):160-7. PMC: 212855. DOI: 10.1128/jb.167.1.160-167.1986. View

20.

Lopilato J, Bortner S, Beckwith J . Mutations in a new chromosomal gene of Escherichia coli K-12, pcnB, reduce plasmid copy number of pBR322 and its derivatives. Mol Gen Genet. 1986; 205(2):285-90. DOI: 10.1007/BF00430440. View