Development of antibody to human GM3 synthase and immunodetection of the enzyme in human tissues

Polyclonal antibody was raised to a cloned fragment of human GM3 synthase. Affinity purified R27C1 antibody to the tagged recombinant protein inhibited GM3 synthase activity in human liver and HL-60 cells in a dose-dependent manner. However, the R27C1 antibody did not affect liver sialyltransferase...

Full description

Saved in:

Bibliographic Details
Published in:	Biochemistry (Moscow) Vol. 69; no. 3; pp. 275 - 280
Main Authors:	Golovanova, N K, Samovilova, N N, Gracheva, E V, Peklo, M M, Vlasik, T N, Sobolev, A Yu, Jurchenko, Yu V, Prokazova, N V
Format:	Journal Article
Language:	English
Published:	United States Springer 01-03-2004 Springer Nature B.V
Subjects:	alpha-Fetoproteins - chemistry Animals Antibodies Antibodies - immunology Antibodies - pharmacology Aorta - enzymology Aorta - immunology Aorta - pathology Arteriosclerosis - enzymology Arteriosclerosis - immunology Arteriosclerosis - pathology Asialoglycoproteins - chemistry Brain - enzymology Brain - immunology Fetuins HL-60 Cells Humans Kinetics Liver Liver - enzymology Liver - immunology Molecular Weight Organ Specificity - immunology Proteins Rabbits Recombinant Proteins - chemistry Recombinant Proteins - genetics Recombinant Proteins - immunology Recombinant Proteins - metabolism Rodents Sialyltransferases - antagonists & inhibitors Sialyltransferases - chemistry Sialyltransferases - genetics Sialyltransferases - immunology Sialyltransferases - metabolism Tunica Intima - enzymology Tunica Intima - immunology Tunica Intima - pathology Viral antibodies
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	Polyclonal antibody was raised to a cloned fragment of human GM3 synthase. Affinity purified R27C1 antibody to the tagged recombinant protein inhibited GM3 synthase activity in human liver and HL-60 cells in a dose-dependent manner. However, the R27C1 antibody did not affect liver sialyltransferase activity towards asialofetuin. We are the first to measure GM3 synthase activity in human liver (194 +/- 60 pmol NeuAc/h per mg protein), which was about 10-fold lower than in phorbol myristate acetate-stimulated HL-60 cells (1353 +/- 573 pmol NeuAc/h per mg protein). On immunoblotting the R27C1 antibody recognized a common protein band in a number of human tissues (liver, brain, atherosclerotic aortic intima, HL-60 cells) with molecular mass of about 60 kD, which is similar to that of the purified GM3 synthase from rat liver. In human liver and aortic intima, the 60-kD band was almost a single band, which makes possible the use of the R27C1 antibody for immunohistochemical studies in these tissues.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0006-2979 1608-3040
DOI:	10.1023/B:BIRY.0000022057.62747.16