Mammalian sulphatases (EC 3.1.6) are a family of enzymes that have a high degree of similarity in amino acid sequence, structure and catalytic mechanism. IDS (iduronate-2-sulphatase; EC 3.1.6.13) is a lysosomal exo-sulphatase that belongs to this protein family and is involved in the degradation of the glycosaminoglycans heparan sulphate and dermatan sulphate. An IDS deficiency causes the lysosomal storage disorder MPS II (mucopolysaccharidosis type II). To examine the structural alterations in heat-denatured and mutant IDS, a panel of four monoclonal antibodies was raised to the denatured protein and used as probes of protein conformation. The linear sequence epitope reactivity of a polyclonal antibody raised against the native protein and the monoclonal antibodies were defined and mapped to distinct regions on the IDS protein. The antigenicity of native IDS was higher in regions without glycosylation, but reactivity was not restricted to protein surface epitopes. One monoclonal epitope was relatively surface accessible and in close proximity to an N-linked glycosylation site, while three others required additional thermal energy to expose the epitopes. The monoclonal antibodies demonstrated the capacity to differentiate progressive structural changes in IDS and could be used to characterize the severity of MPS type II in patients based on variable denatured microstates.
Skip Nav Destination
Article navigation
March 2005
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
February 22 2005
Analysis of normal and mutant iduronate-2-sulphatase conformation
Emma PARKINSON-LAWRENCE;
Emma PARKINSON-LAWRENCE
*Lysosomal Diseases Research Unit, Department of Genetic Medicine, Women's and Children's Hospital, 72 King William Rd, North Adelaide, South Australia 5006, Australia
Search for other works by this author on:
Christopher TURNER;
Christopher TURNER
*Lysosomal Diseases Research Unit, Department of Genetic Medicine, Women's and Children's Hospital, 72 King William Rd, North Adelaide, South Australia 5006, Australia
Search for other works by this author on:
John HOPWOOD;
John HOPWOOD
*Lysosomal Diseases Research Unit, Department of Genetic Medicine, Women's and Children's Hospital, 72 King William Rd, North Adelaide, South Australia 5006, Australia
†Department of Paediatrics, University of Adelaide, Adelaide, South Australia 5005, Australia
Search for other works by this author on:
Doug BROOKS
Doug BROOKS
1
*Lysosomal Diseases Research Unit, Department of Genetic Medicine, Women's and Children's Hospital, 72 King William Rd, North Adelaide, South Australia 5006, Australia
†Department of Paediatrics, University of Adelaide, Adelaide, South Australia 5005, Australia
1To whom correspondence should be addressed, at Lysosomal Diseases Research Unit, Department of Genetic Medicine, Women's and Children's Hospital (email douglas.brooks@adelaide.edu.au).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
May 05 2004
Revision Received:
October 14 2004
Accepted:
October 25 2004
Accepted Manuscript online:
October 25 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2005
Biochem J (2005) 386 (2): 395–400.
Article history
Received:
May 05 2004
Revision Received:
October 14 2004
Accepted:
October 25 2004
Accepted Manuscript online:
October 25 2004
Citation
Emma PARKINSON-LAWRENCE, Christopher TURNER, John HOPWOOD, Doug BROOKS; Analysis of normal and mutant iduronate-2-sulphatase conformation. Biochem J 1 March 2005; 386 (2): 395–400. doi: https://doi.org/10.1042/BJ20040739
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.