Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Bovine kidney beta-mannosidase: purification and characterization

B L Sopher1, C E Traviss, K T Cavanagh

  • 1Department of Pathology, Michigan State University, East Lansing 48824.

The Biochemical Journal
|January 15, 1993
PubMed
Summary

Researchers purified lysosomal beta-mannosidase from bovine kidney, identifying three related peptides crucial for enzyme activity. These peptides were absent in kidneys from beta-mannosidosis affected calves.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

A novel DFNB1 deletion allele supports the existence of a distant cis-regulatory region that controls GJB2 and GJB6 expression.

Clinical genetics·2010
Same author

Genetic mutation in pontocerebellar hypoplasia.

Clinical genetics·2009
Same author

Depressive symptoms in mid-pregnancy, lifetime stressors and the 5-HTTLPR genotype.

Genes, brain, and behavior·2006
Same author

Association and linkage of alpha-2A adrenergic receptor gene polymorphisms with childhood ADHD.

Molecular psychiatry·2004
Same author

Mutations in the gamma-actin gene (ACTG1) are associated with dominant progressive deafness (DFNA20/26).

American journal of human genetics·2003
Same author

Polyglutamine-expanded ataxin-7 antagonizes CRX function and induces cone-rod dystrophy in a mouse model of SCA7.

Neuron·2001

Area of Science:

  • Biochemistry
  • Enzymology
  • Molecular Biology

Background:

  • Lysosomal beta-mannosidase deficiency causes beta-mannosidosis, a genetic disorder.
  • Understanding the enzyme's structure is key to developing potential therapies.

Purpose of the Study:

  • To purify and characterize lysosomal beta-mannosidase from bovine kidney.
  • To investigate the protein composition and immunological properties of the purified enzyme.

Main Methods:

  • Enzyme purification using concanavalin A-Sepharose, immunoaffinity, and HPLC cation-exchange chromatography.
  • Analysis of purified enzyme by SDS/PAGE, Coomassie Blue, and silver staining.
  • Immunological characterization using monoclonal and polyclonal antibodies.
  • N-linked carbohydrate removal and subsequent molecular size analysis.

Related Experiment Videos

Main Results:

  • Lysosomal beta-mannosidase was purified 160,000-fold with a 24% yield.
  • The purified enzyme preparation contained three immunologically related peptides (84, 100, and 110 kDa) associated with enzyme activity.
  • Removal of N-linked carbohydrates reduced the molecular sizes of these peptides.
  • These peptides were undetectable in bovine kidneys from calves with beta-mannosidosis.

Conclusions:

  • The study successfully purified and characterized bovine kidney lysosomal beta-mannosidase.
  • The identified peptides are integral components of the enzyme and are absent in beta-mannosidosis.
  • This provides a foundation for further research into the molecular basis of beta-mannosidosis.