Selective extraction of subunit D of the Na(+)-translocating methyltransferase and subunit c of the A(1)A(0) ATPase from the cytoplasmic membrane of methanogenic archaea by chloroform/methanol and characterization of subunit c of Methanothermobacter thermoautotrophicus as a 16-kDa proteolipid.
Chloroform/methanol was applied to cytoplasmic membranes of the thermophilic methanogens Methanothermobacter thermoautotrophicus and Methanothermobacter marburgensis as well as to the mesophile Methanosarcina mazei Gö1. In any case, the chloroform/methanol extraction yielded only two proteins, subunit D (MtrD) of the Na(+)-translocating methyltetrahydromethanopterin:coenzyme M methyltransferase and the proteolipid of the A(1)A(0) ATPase. Both polypeptides are assumed to be directly involved in ion translocation in their respective enzymes, but have not been studied in detail due to lack of simple isolation procedures. The rapid and selective isolation by chloroform/methanol offers a new way to obtain the large quantities of material required for biochemical analyses. As a first result, molecular and biochemical data suggest that the proteolipid from M. thermoautotrophicus is a duplication of the 8-kDa proteolipid usually present in other archaea, but it retained the conserved glutamate involved in proton translocation in every copy. This is the first 16-kDa proteolipid found in archaea.
Ruppert
C
C
Lehrstuhl für Mikrobiologie der Ludwig-Maxilians-Universität München, Germany.
Schmid
R
R
Hedderich
R
R
Müller
V
V
D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry / Q000302:isolation & purification; D002725:Chloroform; D008565:Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification; D017013:Methanobacteriaceae / Q000737:chemistry / Q000145:classification / Q000201:enzymology*; D000432:Methanol; D017020:Methanosarcina / Q000737:chemistry / Q000145:classification / Q000201:enzymology*; D008780:Methyltransferases / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D021122:Protein Subunits; D011510:Proteolipids / Q000737:chemistry*
D019843:Archaeal Proteins; D008565:Membrane Proteins; D021122:Protein Subunits; D011510:Proteolipids; D002725:Chloroform; D008780:Methyltransferases; C048755:tetrahydromethanopterin S-methyltransferase; D000251:Adenosine Triphosphatases; D000432:Methanol