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Coiled-coil structure of group A streptococcal M proteins. Different temperature stability of class A and C proteins by hydrophobic-nonhydrophobic amino acid substitutions at heptad positions a and d

Cedervall, T LU ; Johansson, M U LU and Akerström, B LU (1997) In Biochemistry 36(16). p.4987-4994
Abstract

M proteins and M-like proteins, expressed on the surface of group A streptococci and binding to human plasma proteins, can be divided into two classes, A and C, depending on the structure of the central repeated regions. The class C proteins have been shown to be dimers with a coiled-coil structure. In this work, we have compared the structure and binding of a class A protein, Mrp4, and a class C protein, Arp4, expressed by the same bacterial strain. Circular dichroism spectra, gel filtration, and binding assays showed that both proteins had a coiled-coil dimer configuration and a high-affinity binding at 20 degrees C. However, striking differences were seen at 37 degrees C. The class A protein, Mrp4, was still a coiled-coil dimer with... (More)

M proteins and M-like proteins, expressed on the surface of group A streptococci and binding to human plasma proteins, can be divided into two classes, A and C, depending on the structure of the central repeated regions. The class C proteins have been shown to be dimers with a coiled-coil structure. In this work, we have compared the structure and binding of a class A protein, Mrp4, and a class C protein, Arp4, expressed by the same bacterial strain. Circular dichroism spectra, gel filtration, and binding assays showed that both proteins had a coiled-coil dimer configuration and a high-affinity binding at 20 degrees C. However, striking differences were seen at 37 degrees C. The class A protein, Mrp4, was still a coiled-coil dimer with high affinity binding activity, whereas the class C protein, Arp4, had lost both the coiled-coil structure and binding activity. Raising the temperature even higher, Mrp4 retained the coiled-coil structure up to 70-90 degrees C. Furthermore, a recombinant protein, Mrp(C), in which the A-repeats of Mrp4 were replaced by the C-repeats of Arp4, lost its coiled-coil structure and fibrinogen-binding around 40-45 degrees C. These results suggest a high thermal stability of class A proteins and a low stability of class C proteins and that the structural basis for this can be found partly in the A- and C-repeats. Analysis of the amino acid sequences of the A- and C-repeats, revealed a large difference, 87% and 45%, respectively, in the content of hydrophobic amino acid residues in the positions regarded as important for the formation of the coiled-coil structure. In particular, several alanine residues in the A-repeats were replaced by serine residues in the C-repeats. Our results suggest that important structural and functional changes within the M protein family have evolved by specific hydrophobic-nonhydrophobic amino acid replacements.

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published
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keywords
Amino Acid Sequence, Amino Acids/analysis, Antigens, Bacterial/chemistry, Antigens, Surface/chemistry, Bacterial Outer Membrane Proteins, Bacterial Proteins/chemistry, Carrier Proteins/chemistry, Chromatography, Gel, Circular Dichroism, Fibrinogen/metabolism, Hot Temperature, Humans, Molecular Sequence Data, Mutagenesis, Site-Directed, Protein Conformation, Protein Structure, Secondary, Recombinant Proteins/metabolism, Streptococcus pyogenes/chemistry, Structure-Activity Relationship
in
Biochemistry
volume
36
issue
16
pages
4987 - 4994
publisher
The American Chemical Society (ACS)
external identifiers
  • scopus:0030887662
ISSN
0006-2960
DOI
10.1021/bi962971q
language
English
LU publication?
yes
id
ba53f522-1fc5-490c-af2f-df165ba3f46b
date added to LUP
2019-05-22 10:18:07
date last changed
2019-11-05 05:19:37
@article{ba53f522-1fc5-490c-af2f-df165ba3f46b,
  abstract     = {<p>M proteins and M-like proteins, expressed on the surface of group A streptococci and binding to human plasma proteins, can be divided into two classes, A and C, depending on the structure of the central repeated regions. The class C proteins have been shown to be dimers with a coiled-coil structure. In this work, we have compared the structure and binding of a class A protein, Mrp4, and a class C protein, Arp4, expressed by the same bacterial strain. Circular dichroism spectra, gel filtration, and binding assays showed that both proteins had a coiled-coil dimer configuration and a high-affinity binding at 20 degrees C. However, striking differences were seen at 37 degrees C. The class A protein, Mrp4, was still a coiled-coil dimer with high affinity binding activity, whereas the class C protein, Arp4, had lost both the coiled-coil structure and binding activity. Raising the temperature even higher, Mrp4 retained the coiled-coil structure up to 70-90 degrees C. Furthermore, a recombinant protein, Mrp(C), in which the A-repeats of Mrp4 were replaced by the C-repeats of Arp4, lost its coiled-coil structure and fibrinogen-binding around 40-45 degrees C. These results suggest a high thermal stability of class A proteins and a low stability of class C proteins and that the structural basis for this can be found partly in the A- and C-repeats. Analysis of the amino acid sequences of the A- and C-repeats, revealed a large difference, 87% and 45%, respectively, in the content of hydrophobic amino acid residues in the positions regarded as important for the formation of the coiled-coil structure. In particular, several alanine residues in the A-repeats were replaced by serine residues in the C-repeats. Our results suggest that important structural and functional changes within the M protein family have evolved by specific hydrophobic-nonhydrophobic amino acid replacements.</p>},
  author       = {Cedervall, T and Johansson, M U and Akerström, B},
  issn         = {0006-2960},
  keyword      = {Amino Acid Sequence,Amino Acids/analysis,Antigens, Bacterial/chemistry,Antigens, Surface/chemistry,Bacterial Outer Membrane Proteins,Bacterial Proteins/chemistry,Carrier Proteins/chemistry,Chromatography, Gel,Circular Dichroism,Fibrinogen/metabolism,Hot Temperature,Humans,Molecular Sequence Data,Mutagenesis, Site-Directed,Protein Conformation,Protein Structure, Secondary,Recombinant Proteins/metabolism,Streptococcus pyogenes/chemistry,Structure-Activity Relationship},
  language     = {eng},
  month        = {04},
  number       = {16},
  pages        = {4987--4994},
  publisher    = {The American Chemical Society (ACS)},
  series       = {Biochemistry},
  title        = {Coiled-coil structure of group A streptococcal M proteins. Different temperature stability of class A and C proteins by hydrophobic-nonhydrophobic amino acid substitutions at heptad positions a and d},
  url          = {http://dx.doi.org/10.1021/bi962971q},
  volume       = {36},
  year         = {1997},
}