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The quaternary structure and activity of newly purified fatty acid synthetase from the Harderian gland of guinea-pig
Journal article   Peer reviewed

The quaternary structure and activity of newly purified fatty acid synthetase from the Harderian gland of guinea-pig

T Kitamoto, M Nishigai, A Ikai, K Ohashi and Y Seyama
Biochimica et biophysica acta. Protein structure and molecular enzymology, Vol.827(2), pp.164-173
02/04/1985
DOI: 10.1016/0167-4838(85)90086-x
PMID: 3967036

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Abstract

Fatty acid synthetase was isolated from the Harderian gland of guinea-pig. The fatty acids synthesized by the purified enzyme were analyzed by mass fragmentography. The purified enzyme had an inherent capacity to utilize methylmalonyl-CoA and synthesize methyl-branched fatty acids. Physicochemical studies indicated that an active enzyme was a dimer, consisted of two subunits of Mr = 2.5 X 10(5). The negatively stained enzyme had an electron micrographic image of an ellipsoidal contour with a continuous middle cleft along the major axis. The major and minor axes were approximately equal to 220 and 150 A, respectively. In a dimer, the subunit had a rod-like structure about 220 A long and 50 A wide. The enzyme was inactivated and dissociated into subunits by incubation at 0 degree C. The inactivated enzyme was fully reactivated by raising the temperature of the solution. The relationship between the quaternary structure of the enzyme and the occurrence of enzymatic activity was studied by high-performance liquid chromatography. Neither active monomers nor inactive dimers were found in inactivation and reactivation processes. The initial velocity of reactivation was proportional to the enzyme concentration over a concentration range of 160-800 micrograms/ml, indicating that the rate-determining step in the reactivation reaction was unimolecular.
Fatty Acid Synthases - antagonists & inhibitors Fatty Acid Synthases - isolation & purification Animals Guinea Pigs Harderian Gland - enzymology Fatty Acid Synthases - metabolism Lacrimal Apparatus - enzymology Amino Acids - analysis Protein Conformation Microscopy, Electron Circular Dichroism Macromolecular Substances

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