Cytochrome b-562 from Acinetobacter calcoaceticus L.M.D. 79.41. Its characteristics and role as electron acceptor for quinoprotein glucose dehydrogenase.
A soluble cytochrome b was purified from Acinetobacter calcoaceticus L.M.D. 79.41. On the basis of the alpha-band maximum of a reduced preparation, measured at 25 degrees C, it is designated as cytochrome b-562. This cytochrome is a basic monomeric protein (pI 10.2; Mr 18,000), containing one protohaem group per molecule. The reduced form, at 25 degrees C, showed absorption bands at 428, 532 and 562 nm. At 77 K the alpha-band shifted to 560 nm (with a shoulder at 558 nm). The reduced cytochrome did not react with CO. Cytochrome b-562 is most probably (loosely) attached to the outside of the cytoplasmic membrane, since substantial amounts of it, equimolar to quinoprotein glucose dehydrogenase (GDH), were present in the culture medium when cells were grown in the presence of low concentrations of Triton X-100. The midpoint potential at pH 7.0 was found to be +170 mV, a value that was lowered to +145 mV by the presence of GDH. Since the GDH was shown to have a midpoint potential of +50 mV, cytochrome b-562 could function as the natural primary electron acceptor. Arguments to substantiate this view and to propose a role of ubiquinone-9 as electron acceptor for cytochrome b-562 are presented.
Dokter
P
P
Delft University of Technology, Laboratory of Microbiology and Enzymology, The Netherlands.
van Wielink
J E
JE
van Kleef
M A
MA
Duine
J A
JA
D000150:Acinetobacter / Q000235:genetics / Q000378:metabolism*; D002237:Carbohydrate Dehydrogenases / Q000378:metabolism*; D003573:Cytochrome b Group / Q000302:isolation & purification / Q000378:metabolism*; D004579:Electron Transport; D029968:Escherichia coli Proteins; D005947:Glucose / Q000378:metabolism; D005948:Glucose Dehydrogenases / Q000378:metabolism*; D006418:Heme / Q000032:analysis; D008970:Molecular Weight; D009154:Mutation; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D011199:Potentiometry; D013053:Spectrophotometry; D014451:Ubiquinone / Q000378:metabolism
D003573:Cytochrome b Group; D029968:Escherichia coli Proteins; D014451:Ubiquinone; D006418:Heme; C017850:cytochrome b562, E coli; D002237:Carbohydrate Dehydrogenases; D005948:Glucose Dehydrogenases; C033955:glucose dehydrogenase (pyrroloquinoline-quinone); D005947:Glucose