NDLI: Partial purification and characterization of sphingosine N-acyltransferase (ceramide synthase) from bovine liver mitochondrion-rich fraction

Content Provider	Springer Nature Link
Author	Shime, Hiroshi Soeda, Shinji Sakamoto, Masahiko Kouchi, Takeshi Kowakame, Takashi Kihara, Taro
Copyright Year	1998
Abstract	Sphingosine N-acyltransferase (ceramide synthase, E.C. 2.3.1.24) was solubilized from bovine liver mitochondrion-rich fraction with n-ocytl β-d-thioglucoside as the detergent and partially purified by sequential chromatography on columns of DE-32, shingosine affinity, and Sepharose CL-6B. The partially purified preparation migrated on SDS-polyacrylamide gel electrophoresis as two major protein bands of 62 and 72 kDa. The molecular mass of the enzyme estimated by gel filtration was 240–260 kDa, suggesting that the partially purified enzyme is present in a subunit form or simply has an aggregative nature. The specific activity of the final preparation for the condensation of sphingosine with stearoyl-CoA increased by 98.7-fold compared with the starting material. The optimal pH value for the ceramide synthesis was 7.5. The partially purified enzyme had an apparent K $_{m}$ of 146 μM and a V $_{max}$ of 11.1 nmol/min/mg protein for stearoyl-CoA. The K $_{m}$ and V $_{max}$ values toward sphingosine were 171 μM and 11.3 nmol/min/mg protein, respectively. Interestingly, sphinganine was also a good substrate for this enzyme, and the K $_{m}$ and V $_{max}$ values were 144 μM and 8.5 nmol/min/mg protein, respectively.
Starting Page	601
Ending Page	605
Page Count	5
File Format	PDF
ISSN	00244201
Journal	Lipids
Volume Number	33
Issue Number	6
e-ISSN	15589307
Language	English
Publisher	Springer-Verlag
Publisher Date	1998-01-01
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Nutrition Bioorganic Chemistry Medicinal Chemistry Medical Biochemistry Biochemistry Microbial Genetics and Genomics
Content Type	Text
Resource Type	Article
Subject	Cell Biology Organic Chemistry Biochemistry

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