Metal binding is critical for the folding and function of laminin binding protein, Lmb of Streptococcus agalactiae.

Lmb is a 34 kDa laminin binding surface adhesin of Streptococcus agalactiae. The structure of Lmb reported by us recently has shown that it consists of a metal binding crevice, in which a zinc ion is coordinated to three highly conserved histidines. To elucidate the structural and functional signifi...

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Autores principales:	Preethi Ragunathan, Divya Sridaran, Anja Weigel, Sarah Shabayek, Barbara Spellerberg, Karthe Ponnuraj
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8768567b659342ee82212129f28b1d21
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spelling	oai:doaj.org-article:8768567b659342ee82212129f28b1d212021-11-18T07:40:16ZMetal binding is critical for the folding and function of laminin binding protein, Lmb of Streptococcus agalactiae.1932-620310.1371/journal.pone.0067517https://doaj.org/article/8768567b659342ee82212129f28b1d212013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23826314/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Lmb is a 34 kDa laminin binding surface adhesin of Streptococcus agalactiae. The structure of Lmb reported by us recently has shown that it consists of a metal binding crevice, in which a zinc ion is coordinated to three highly conserved histidines. To elucidate the structural and functional significance of the metal ion in Lmb, these histidines have been mutated to alanine and single, double and triple mutants were generated. These mutations resulted in insolubility of the protein and revealed altered secondary and tertiary structures, as evidenced by circular dichroism and fluorescence spectroscopy studies. The mutations also significantly decreased the binding affinity of Lmb to laminin, implicating the role played by the metal binding residues in maintaining the correct conformation of the protein for its binding to laminin. A highly disordered loop, proposed to be crucial for metal acquisition in homologous structures, was deleted in Lmb by mutation (ΔLmb) and its crystal structure was solved at 2.6 Å. The ΔLmb structure was identical to the native Lmb structure with a bound zinc ion and exhibited laminin binding activity similar to wild type protein, suggesting that the loop might not have an important role in metal acquisition or adhesion in Lmb. Targeted mutations of histidine residues confirmed the importance of the zinc binding crevice for the structure and function of the Lmb adhesin.Preethi RagunathanDivya SridaranAnja WeigelSarah ShabayekBarbara SpellerbergKarthe PonnurajPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 6, p e67517 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Preethi Ragunathan Divya Sridaran Anja Weigel Sarah Shabayek Barbara Spellerberg Karthe Ponnuraj Metal binding is critical for the folding and function of laminin binding protein, Lmb of Streptococcus agalactiae.
description	Lmb is a 34 kDa laminin binding surface adhesin of Streptococcus agalactiae. The structure of Lmb reported by us recently has shown that it consists of a metal binding crevice, in which a zinc ion is coordinated to three highly conserved histidines. To elucidate the structural and functional significance of the metal ion in Lmb, these histidines have been mutated to alanine and single, double and triple mutants were generated. These mutations resulted in insolubility of the protein and revealed altered secondary and tertiary structures, as evidenced by circular dichroism and fluorescence spectroscopy studies. The mutations also significantly decreased the binding affinity of Lmb to laminin, implicating the role played by the metal binding residues in maintaining the correct conformation of the protein for its binding to laminin. A highly disordered loop, proposed to be crucial for metal acquisition in homologous structures, was deleted in Lmb by mutation (ΔLmb) and its crystal structure was solved at 2.6 Å. The ΔLmb structure was identical to the native Lmb structure with a bound zinc ion and exhibited laminin binding activity similar to wild type protein, suggesting that the loop might not have an important role in metal acquisition or adhesion in Lmb. Targeted mutations of histidine residues confirmed the importance of the zinc binding crevice for the structure and function of the Lmb adhesin.
format	article
author	Preethi Ragunathan Divya Sridaran Anja Weigel Sarah Shabayek Barbara Spellerberg Karthe Ponnuraj
author_facet	Preethi Ragunathan Divya Sridaran Anja Weigel Sarah Shabayek Barbara Spellerberg Karthe Ponnuraj
author_sort	Preethi Ragunathan
title	Metal binding is critical for the folding and function of laminin binding protein, Lmb of Streptococcus agalactiae.
title_short	Metal binding is critical for the folding and function of laminin binding protein, Lmb of Streptococcus agalactiae.
title_full	Metal binding is critical for the folding and function of laminin binding protein, Lmb of Streptococcus agalactiae.
title_fullStr	Metal binding is critical for the folding and function of laminin binding protein, Lmb of Streptococcus agalactiae.
title_full_unstemmed	Metal binding is critical for the folding and function of laminin binding protein, Lmb of Streptococcus agalactiae.
title_sort	metal binding is critical for the folding and function of laminin binding protein, lmb of streptococcus agalactiae.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/8768567b659342ee82212129f28b1d21
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Metal binding is critical for the folding and function of laminin binding protein, Lmb of Streptococcus agalactiae.

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