The sodium ion translocating oxalacetate decarboxylase of Klebsiella pneumoniae. Sequence of the biotin-containing alpha-subunit and relationship to other biotin-containing enzymes.
The gene encoding the alpha-subunit of the Na+ pump oxalacetate decarboxylase of Klebsiella pneumoniae was cloned and sequenced. The deduced primary structure of the protein was confirmed by protein sequencing of about 30% of the polypeptide chain. The gene has a GC content of 67% and codes for 596 amino acids. The N-terminal methionine is removed in the mature protein which has a calculated molecular mass of 63,600 daltons. The protein consists of two different domains that are connected by a stretch of amino acid residues susceptible to proteolytic cleavage. Limited proteolysis of the native enzyme with trypsin produced fragments of about 51 kDa and 10.2 kDa, the latter of which started with valine 491 and contained the biotin prosthetic group. Peptide sequencing indicated binding of the biotin prosthetic group to lysine 561, 35 residues from the C terminus. The decarboxylase contains an extended alanine- and proline-rich region (positions 502-532) on the N-terminal side of the 10.2-kDa biotin domain. This sequence includes a total of 16 alanine and 9 proline residues.
Schwarz
E
E
Max-Planck-Institut für Biochemie, Martinsried, Federal Republic of Germany.
Oesterhelt
D
D
Reinke
H
H
Beyreuther
K
K
Dimroth
P
P
D000409:Alanine; D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D001710:Biotin / Q000378:metabolism*; D002262:Carboxy-Lyases / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D007473:Ion Channels / Q000378:metabolism; D007711:Klebsiella pneumoniae / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000378:metabolism; D011392:Proline; D012697:Serine Endopeptidases / Q000378:metabolism; D012964:Sodium / Q000378:metabolism*; D014357:Trypsin / Q000378:metabolism
D007473:Ion Channels; D010446:Peptide Fragments; D001710:Biotin; D011392:Proline; D012964:Sodium; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D014357:Trypsin; D002262:Carboxy-Lyases; C021916:oxaloacetate decarboxylase; D000409:Alanine