Skip Navigation

This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Request Permissions
Google Scholar
Right arrow Articles by SAKURAGAWA, N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by SAKURAGAWA, N.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

J. Biochem, 1982, Vol. 92, No. 3 637-646
© 1982 Japanese Biochemical Society

research-article

Acid Sphingomyelinase of Human Placenta: Purification, Properties, and 125Iodine Labeling1

Norio SAKURAGAWA2

Developmental and Metabolic Neurology Branch, National Institute of Neurological and Communicative Diseases and Stroke Bethesda, Maryland, 20014, U.S.A.

Human placental acid sphingomyelinase was highly purified in the presence of Triton X-100. DEAE-Sephacel chromatography and chromatofocusing were the most effective steps in the purification procedure. Enzyme purification was 380,000 nmol/mg protein/h. Characterization and radioiodination were carried out with the chromatofocusing fraction containing highly purified enzyme. The purified enzyme contained no activity of eleven other lysosomal hydrolases but hydrolyzed bis-p-nitrophenyl phosphate slowly compared with (14C)sphingomyelin and chromogenic substrates. SDS-gel electrophoresis revealed two distinct protein bands with molecular weights of 70,500 and 39,800. This enzyme had a molecular weight of 200,000 as determined by analytical gel filtration. The pH optimum was 5.0 and Km was 52.6 × 10–5 M for [14C)sphingomyelin.

Highly purified sphingomyelinase was labeled with 125iodine by the use of Enzymobeads. Labeled sphingomyelinase preparation was rapidly cleared from blood with t1/2 of 1 min. It was absorbed mostly into the liver and presumably largely excreted from there. This labeled enzyme may be useful in metabolic studies in normal animals and animal models of genetic lysosomal storage disorders.

1 This study was supported in part by a Grant-in-Aid for New Drug Development from the Ministry of Health and Welfare, Japan.

2 Present address: Division of Child Neurology, National Center for Nervous, Mental and Muscular Diseases, 2620, Ogawa-Higashi-Machi, Kodaira, Tokyo 187.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 FASEB J.Home page
E. L. Smith and E. H. Schuchman
The unexpected role of acid sphingomyelinase in cell death and the pathophysiology of common diseases
FASEB J, October 1, 2008; 22(10): 3419 - 3431.
[Abstract] [Full Text] [PDF]

Home page
 J. Lipid Res.Home page
Y. Cheng, A. Nilsson, E. Tomquist, and R.-D. Duan
Purification, characterization, and expression of rat intestinal alkaline sphingomyelinase
J. Lipid Res., February 1, 2002; 43(2): 316 - 324.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.