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Single Particle Reconstruction of Human Fatty Acid Synthase

Published online by Cambridge University Press:  02 July 2020

J. Brink ,
S.J. Ludtke ,
C.-Y. Yang ,
Z.-W. Gu ,
S. Wakil  and
W. Chiu
J. Brink
Affiliation:
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, HoustonTX77030
S.J. Ludtke
Affiliation:
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, HoustonTX77030
C.-Y. Yang
Affiliation:
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, HoustonTX77030
Z.-W. Gu
Affiliation:
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, HoustonTX77030
S. Wakil
Affiliation:
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, HoustonTX77030
W. Chiu
Affiliation:
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, HoustonTX77030
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Extract

Fatty acid synthase (FAS) is the enzyme responsible for de novo synthesis of fatty acids from acetyl-CoA, malonyl-CoA and NADPH. FAS (550 kDa) is a homodimer of two multifunctional polypeptides, each with seven distinct catalytic activities and a site for the prosthetic group, 4’- phosphopantetheine, acyl carrier protein (ACP). These domains are organized from the N- to the C-terminus as follows: keto acylsynthase, acetyl/malonyl transacetylase, dehydratase, the interdomain, enoyl reductase, ketoreductase, ACP and thioesterase. The two polypeptides are held together through the interdomain and oriented in an anti-parallel manner, each contributing complementary half sites and giving rise to two independently active centers for palmitate synthesis. Interest in FAS arises from its involvement in human disorders, such as obesity, hyperlipidemia and carcinogenesis.

Human FAS purified from a breast cancer cell line, ZR75-1, was vitrified at 50-70 μg/ml on holey grids in the presence of NADPH and acetyl-CoA and kept at -166°C in a Gatan 626 cryo-holder.

Type
Electron Cryomicroscopy of Macromolecules
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 270 - 271
Copyright
Copyright © Microscopy Society of America

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References

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5. This research has been supported by grants from NIH (RR02250 and GM19091) and from NCS (MCB990021N).Google Scholar