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  Domain IV of mouse laminin beta 1 and beta 2 chains - Structure, glycosaminoglycan modification and immunochemical analysis of tissue contents

Sasaki, T., Mann, K., Miner, J. H., Miosge, N., & Timpl, R. (2002). Domain IV of mouse laminin beta 1 and beta 2 chains - Structure, glycosaminoglycan modification and immunochemical analysis of tissue contents. European Journal of Biochemistry, 269(2), 431-442.

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Genre: Journal Article
Alternative Title : Eur. J. Biochem.

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 Creators:
Sasaki, T.1, Author           
Mann, K.2, 3, Author           
Miner, J. H., Author
Miosge, N., Author
Timpl, R.1, Author           
Affiliations:
1Former Research Groups, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565145              
2Mann, Matthias / Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565159              
3Huber, Robert / Structure Research, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565155              

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Free keywords: basement membranes; chondroitin sulfate; laminin domain; radioimmunoassay; recombinant production
 Abstract: Domain IV, consisting of about 230 residues, represents a particular protein module so far found only in laminin beta1 and beta2 chains. Both domains were obtained by recombinant production in mammalian cells. They showed a globular structure, as expected from electron microscopic examination of laminins. Fragment beta1IV was obtained as a monomer and a disulfide-bonded dimer, and both were modified to approximate to50% by a single chondroitin sulfate chain attached to Ser721 of an SGD consensus sequence. Dimerization is caused by an odd number of cysteines, with three of them having a partial thiol character. Whether both modifications also occur in tissue forms of laminin remains to be established. Fragment beta21V was only obtained as a monomer, as it lacked one crucial cysteine and the SGD sequence. It required, however, the presence of two adjacent LE modules for proper folding. Polyclonal antibodies raised against both fragments showed no cross-reaction with each other and allowed establishment of beta chain-specific radioimmunoassays and light and electron microscopic immunostaining of tissues. This demonstrated a 5- 25-fold lower content of beta2 compared with beta1 chains in various tissue extracts of adult mice. Tissues derived from beta2-deficient mice failed to react with the beta2-specific antibodies but showed a twofold higher content of beta1 than heterozygotes. The antibodies to beta2 showed broader tissue staining than reported previously, including in particular a distinct reaction with the extra-synaptic endomysium of skeletal muscle. Immunogold staining localized both beta chains primarily to basement membranes of kidney, muscle and various other tissues.

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Language(s): eng - English
 Dates: 2002-01
 Publication Status: Issued
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 39028
ISI: 000173651700004
 Degree: -

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Title: European Journal of Biochemistry
  Alternative Title : Eur. J. Biochem.
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 269 (2) Sequence Number: - Start / End Page: 431 - 442 Identifier: ISSN: 0014-2956