Enzymatic synthesis of 14C-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase: detection of endoglycoceramidase activity in a seaflower.

Enzymatic synthesis of 14C-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase: detection of endoglycoceramidase activity in a seaflower. Research Abstract Details 

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Enzymatic synthesis of 14C-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase: detection of endoglycoceramidase activity in a seaflower. Abstract Text:

S Mitsutake,K Kita,T Nakagawa,M Ito,

This paper describes the synthesis of 14C-labeled glycosphingolipids using the reverse hydrolysis reaction (condensation) of sphingolipid ceramide N-deacylase. It was found that 50-70% of 14C-fatty acids were incorporated into various lyso-glycosphingolipids when a mixture of lyso-glycosphingolipids and fatty acids was incubated at 37 degrees C with 1 mU of the enzyme for 20 h in 1 ml of 25 mM phosphate buffer, pH 6.0-7.0, containing 0-0.1% Triton X-100. The optimum concentration of lyso-glycosphingolipids was 100-400 microM depending on the species of lyso-form when [14C]stearic acid was used at the concentration of 100 microM. Free 14C-fatty acids and lyso-glycosphingolipids were separated from the synthesized 14C-glycosphingolipids by using a Sep-Pak Plus Silica and a Sep-Pak CM or a QMA cartridge, respectively. After treatment of 14C-glycosphingolipids with endoglycoceramidase or sphingolipid ceramide N-deacylase, digestion products were clearly separated from the parent glycosphingolipids on TLC and determined using an image analyzer with a sensitivity 100 times higher than that using non-radiolabeled substrates. Using this method, we found endoglycoceramidase activity in a seaflower, Condylactis sp., for the first time.

Enzymatic synthesis of 14C-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase: detection of endoglycoceramidase activity in a seaflower. Publishing Authors By Initials

S Mitsutake,K Kita,T Nakagawa,M Ito,

For similar animals: invertebrates: cnidaria: anthozoa: sea anemones research abstracts see: animals: invertebrates: cnidaria: anthozoa: sea anemones research

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Enzymatic synthesis of 14C-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase: detection of endoglycoceramidase activity in a seaflower. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of biochemistry

VOLUME: 123

Page Numbers: 859-63

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: May

YEAR: 1998

Enzymatic synthesis of 14C-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase: detection of endoglycoceramidase activity in a seaflower. Information

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LANGUAGE: eng

NlmUniqueID: 376600

Enzymatic synthesis of 14C-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase: detection of endoglycoceramidase activity in a seaflower. Keywords Mesh Terms:

KEYWORDS: Sea Anemones

MESH TERMS: enzymology

Chemical & Substance for Abstract: Enzymatic synthesis of 14C-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase: detection of endoglycoceramidase activity in a seaflower. Information

Substance Name: sphingolipid ceramide N-deacylase

Registry Number: EC 3.5.1.-

Grant and Affiliation Information for Enzymatic synthesis of 14C-glycosphingolipids by reverse hydrolysis reaction of sphingolipid ceramide N-deacylase: detection of endoglycoceramidase activity in a seaflower.

AFFILIATION: Laboratory of Marine Biochemistry, Faculty of Agriculture, Kyushu University, 6-10-1, Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan.

Country: JAPAN

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MEDLINETA: J Biochem

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