PMID: 1267Dec 1, 1975

Investigations of the structure of 3-methylcrotonyl-CoA carboxylase from Achromobacter

European Journal of Biochemistry
U SchieleF Lynen

Abstract

It was shown by gel electrophoresis in sodium dodecylsulphate solution that 3-methylcrotonyl-CoA carboxylase from Achromobacter IVS is composed of two different subunits with molecular weights of about 78000 and 96000, respectively. The biotin is bound to the heavier subunit. It was previously found that 3-methylcrotonyl-CoA carboxylase contains four biotin molecules per complex. A complex composed of four of each subunit would thus have a molecular weight of about 700000. This is compatible with the molecular weight of 760000 determined earlier by analytical ultracentrifugation. Both subunits were isolated preparatively. As the subunits, unlike the complex, are very sensitive to oxygen, special precautions had to be taken during isolation. The biotin-containing subunit was isolated by chromatography on DEAE-cellulose in 5 M urea. It no longer catalyzed the overall reaction, yet could still carboxylate free biotin. The biotin-free subunit was separated after dissociation of the enzyme by three-days' dialysis at pH 9.8 under nitrogen. On chromatography over a Sepharose-bound avidin column, the biotin-subunit was fixed and the biotin-free subunit was eluted unretarded. The latter subunit showed no enzymic activity. After the addi...Continue Reading

References

Sep 1, 1972·Archives of Biochemistry and Biophysics·C G Kannangara, P K STUMPF
Aug 1, 1969·Proceedings of the National Academy of Sciences of the United States of America·A W AlbertsP R Vagelos
Jan 1, 1970·Annual Review of Biochemistry·J Knappe
Jul 1, 1971·Proceedings of the National Academy of Sciences of the United States of America·R R FallP R Vagelos
Sep 1, 1970·European Journal of Biochemistry·R Apitz-CastroF Lynen
Oct 1, 1966·Journal of Bacteriology·S F Gottlieb

Citations

Jul 19, 2012·World Journal of Microbiology & Biotechnology·César Díaz-PérezJesús Campos-García
Jan 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·E P LauR R Fall
Mar 12, 2004·Annual Review of Plant Physiology and Plant Molecular Biology·Claude AlbanRoland Douce
Dec 1, 1979·CRC Critical Reviews in Biochemistry·F Lynen
Aug 10, 1976·Biochemistry·M L Hector, R R Fall
Jun 1, 1982·Archives of Biochemistry and Biophysics·A R Hunaiti, P E Kolattukudy
Feb 22, 2000·The Journal of Biological Chemistry·A L McKeanE S Wurtele

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