NDLI: Purification, cDNA Cloning, and Expression of UDP-Gal: Glucosylceramide β-1,4-Galactosyltransferase from Rat Brain

Content Provider	Journal of Biological Chemistry (JBC)
Author	Nomura, Tomoko Takizawa, Minoru Aoki, Junken Arai, Hiroyuki Inoue, Keizo Wakisaka, Etsuji Yoshizuka, Naonobu Imokawa, Genji Dohmae, Naoshi Takio, Koji Hattori, Michihiro Matsuo, Noboru
Abstract	Lactosylceramide synthase is an enzyme that catalyzes the transfer of galactose from UDP-Gal to glucosylceramide, and thus participates in the biosynthesis of most glycosphingolipids in mammals. We purified this enzyme over 61,000-fold to near homogeneity with a 29.7% yield from rat brain membrane fractions. The isolation procedure included solubilization with Triton X-100, affinity chromatography on wheat germ agglutinin-agarose and UDP-hexanolamine-agarose, and hydroxylapatite column chromatography, followed by ion exchange chromatography. The final preparation migrated as a broad band with an apparent molecular mass of 61 kDa on SDS-polyacrylamide gel electrophoresis. This apparent molecular mass was reduced to 51 kDa by N-glycanase digestion, suggesting that the enzyme has a glycoprotein nature. The enzyme required Mn2+ for its activity, and glucosylceramide was its preferred substrate. The cDNA for the enzyme was cloned from a rat brain cDNA library. The cDNA insert encoded a polypeptide of 382 amino acid residues, with a molecular weight of 44,776. The polypeptide contained eight putative glycosylation sites and a 20-amino acid residue transmembrane domain at its N terminus. Amino acid sequence homology analysis revealed that this enzyme shared 39% homology with mouse β-1,4-galactosyltransferase (EC 2.4.1.38), which catalyzes the transfer of Gal to β-1,4-GlcNAc in glycoproteins.
Related Links	http://www.jbc.org/content/273/22/13570.abstract
Ending Page	13577
Starting Page	13570
Page Count	8
File Format	HTM / HTML PDF
ISSN	00219258
Journal	Journal of Biological Chemistry (JBC)
Issue Number	22
Volume Number	273
DOI	10.1074/jbc.273.22.13570
e-ISSN	1083351X
Language	English
Publisher	American Society for Biochemistry and Molecular Biology
Publisher Date	1998-05-29
Access Restriction	Open
Subject Keyword	CARBOHYDRATES, LIPIDS, AND OTHER NATURAL PRODUCTS
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Expression Cloning of Rat cDNA Encoding UDP-galactose:GD2 β1,3-galactosyltransferase That Determines the Expression of GD1b/GM1/GA1

Genomic Cloning and Expression of Three Murine UDP-galactose: β-N-Acetylglucosamine β1,3-Galactosyltransferase Genes

Molecular Cloning and Expression of cDNA Encoding the Rat UDP-N-Acetylglucosamine:α-6-D-Mannoside β-1,2-N-Acetylglucosaminyltransferase II

Purification and cDNA Cloning of Porcine Brain GDP-L-Fuc:N-Acetyl-β-D-Glucosaminide α1→6Fucosyltransferase

cDNA Cloning and Expression of a Novel UDP-N-acetyl-d-galactosamine:PolypeptideN-Acetylgalactosaminyltransferase

Purification and cDNA Cloning of a Human UDP-N-acetyl-α- D-galactosamine:polypeptide N-Acetylgalactosaminyltransferase

cDNA Cloning and Expression of Bovine UDP-N-Acetylglucosamine:α1,3-d-Mannoside β1,4-N-Acetylglucosaminyltransferase IV

cDNA Cloning and Expression of a Family of UDP-N-acetyl-dgalactosamine:PolypeptideN-Acetylgalactosaminyltransferase Sequence Homologs fromCaenorhabditis elegans

Purification and Characterization of UDP-glucose:Ceramide Glucosyltransferase from Rat Liver Golgi Membranes

Purification, cDNA Cloning, and Expression of UDP-Gal: Glucosylceramide β-1,4-Galactosyltransferase from Rat Brain

Similar Documents

Expression Cloning of Rat cDNA Encoding UDP-galactose:GD2 β1,3-galactosyltransferase That Determines the Expression of GD1b/GM1/GA1

Genomic Cloning and Expression of Three Murine UDP-galactose: β-N-Acetylglucosamine β1,3-Galactosyltransferase Genes

Molecular Cloning and Expression of cDNA Encoding the Rat UDP-N-Acetylglucosamine:α-6-D-Mannoside β-1,2-N-Acetylglucosaminyltransferase II

Purification and cDNA Cloning of Porcine Brain GDP-L-Fuc:N-Acetyl-β-D-Glucosaminide α1→6Fucosyltransferase

cDNA Cloning and Expression of a Novel UDP-N-acetyl-d-galactosamine:PolypeptideN-Acetylgalactosaminyltransferase

Purification and cDNA Cloning of a Human UDP-N-acetyl-α- D-galactosamine:polypeptide N-Acetylgalactosaminyltransferase

cDNA Cloning and Expression of Bovine UDP-N-Acetylglucosamine:α1,3-d-Mannoside β1,4-N-Acetylglucosaminyltransferase IV

cDNA Cloning and Expression of a Family of UDP-N-acetyl-dgalactosamine:PolypeptideN-Acetylgalactosaminyltransferase Sequence Homologs fromCaenorhabditis elegans

Purification and Characterization of UDP-glucose:Ceramide Glucosyltransferase from Rat Liver Golgi Membranes

Purification, cDNA Cloning, and Expression of UDP-Gal: Glucosylceramide β-1,4-Galactosyltransferase from Rat Brain