Amino terminal sequence of type 3 streptococcal M protein extraction products

1990 ◽  
Vol 55 (10) ◽  
pp. 2562-2567
Author(s):  
Ivan Kluh ◽  
Otto Kühnemund ◽  
Manfred Pavlík ◽  
Ladislav Morávek ◽  
Jiří Havlíček
Keyword(s):  
Protein Extraction ◽  
Peptide Vaccine ◽  
Limited Proteolysis ◽  
Protein Molecule ◽  
M Protein ◽  
Terminal Sequence ◽  
Amino Terminal ◽  
Streptococcal M Protein ◽  
N Terminus ◽  
Type 3

The amino terminal hypervariable part of the M protein molecule was chosen as a basis for the preparation of a synthetic peptide vaccine against group A streptococci. As part of the mapping of various serological types the main products of extraction of type M Streptococcus pyogeneswith limited pepsin (Pep) hydrolysis at pH 5.5 and with phage-associated lysin (PAL) were sequenced. Two entirely different sequences were obtained. The sequence of PAL M3 shows the absence of the α-helical potential in the shorter N-terminal region as is characteristic of the N-terminus of the M protein molecule. The main product of limited hydrolysis Pep M3 (pH 5.5), which shows the presence of the α-helical potential from the very amino terminal residue of its chain, does not involve most likely the proper N-terminus of the M protein. Extraction with pepsin under conditions of very limited proteolysis (pH 5.8) yielded a fragment with N-terminal sequence identical with that of PAL M3 (extracted nonproteolytically).

scholarly journals Multiple binding of type 3 streptococcal M protein to human fibrinogen, albumin and fibronectin

1993 ◽  
Vol 7 (2) ◽  
pp. 135-143 ◽  
Author(s):  
Karl-Hermann Schmidt ◽  
Karlheinz Mann ◽  
Jakki Cooney ◽  
Werner K�hler
Keyword(s):  
M Protein ◽  
Human Fibrinogen ◽  
Streptococcal M Protein ◽  
Multiple Binding ◽  
Type 3

The amino-terminal region of group A streptococcal M protein determines its molecular state of assembly and function

1991 ◽  
Vol 10 (1) ◽  
pp. 49-59 ◽  
Author(s):  
Kiran M. Khandke ◽  
Thomas Fairwell ◽  
Emory H. Braswell ◽  
Belur N. Manjula
Keyword(s):  
Molecular State ◽  
Terminal Region ◽  
M Protein ◽  
Amino Terminal ◽  
Streptococcal M Protein ◽  
Group A ◽  
And Function

scholarly journals Streptococcal M6 protein expressed in Escherichia coli. Localization, purification, and comparison with streptococcal-derived M protein.

1984 ◽  
Vol 159 (4) ◽  
pp. 1083-1095 ◽  
Author(s):  
V A Fischetti ◽  
K F Jones ◽  
B N Manjula ◽  
J R Scott
Keyword(s):  
Apparent Molecular Weight ◽  
M Protein ◽  
Rabbit Serum ◽  
New Host ◽  
E Coli ◽  
Amino Terminal ◽  
Streptococcal M Protein ◽  
Sds Page ◽  
Peptic Digestion ◽  
Amino Terminal Sequence

Type 6 streptococcal M protein produced by E. coli bearing plasmid pJRS42.13 (ColiM6) accumulates in the periplasmic space of this new host. No immunoreactive M protein was found either on the surface of the organism or in the culture medium. The ColiM6 protein was purified from the periplasm and the final preparation consisted of three protein bands of apparent molecular weight 55,000, 57,000, and 59,000. These three bands were identical in migration in SDS PAGE to that of the M protein present in freshly prepared crude periplasm. The amino acid composition of the ColiM6 protein was nearly identical to that of M protein isolated from streptococci with phage lysin (LysM6). Furthermore, except for the amino terminal residue of the LysM6 molecule, the amino terminal sequence of the ColiM6 molecule was identical to those of both LysM6 and M protein released from the streptococcus by limited peptic digestion (PepM6). These results reveal that the molecule produced in the E. coli and transported into the periplasm may be the complete M protein as it exists on the streptococcus. The results also indicate that the systems that process M protein for transport through the cytoplasmic membrane are similar in the streptococcus and E. coli. The purified ColiM6 protein was able to remove opsonic antibodies from both human and rabbit serum, as well as to stimulate the production of opsonic antibodies in rabbits, indicating that the immunodeterminants on this molecule are the same as those found on streptococcal-derived M molecules.

scholarly journals Type-specific immunogenicity of a chemically synthesized peptide fragment of type 5 streptococcal M protein.

1983 ◽  
Vol 158 (5) ◽  
pp. 1727-1732 ◽  
Author(s):  
J B Dale ◽  
J M Seyer ◽  
E H Beachey
Keyword(s):  
Synthetic Peptides ◽  
M Protein ◽  
Antigenic Specificity ◽  
Antigenic Determinants ◽  
Terminal Amino Acid ◽  
Amino Terminal ◽  
Streptococcal M Protein ◽  
Protective Antibodies ◽  
Terminal Amino ◽  
Covalently Linked

We determined the antigenic specificity and protective immunogenicity of two chemically synthesized peptides of type 5 streptococcal M protein. The synthetic peptides, designated S-M5(1-20) and S-M5(20-40), represent the amino-terminal amino acid sequence of the native pepsin-extracted M5 molecule, which is known to contain at least one heart cross-reactive epitope. Initial studies showed that neither of the synthetic peptides was able to bind purified heart-reactive M5 antibodies. In addition, S-M5(1-20), but not S-M5(20-40), contained type-specific antigenic determinants as measured by enzyme-linked immunosorbent inhibition assays. When covalently linked to tetanus toxoid, S-M5(1-20), but not S-M5(20-40), evoked significant levels of type-specific, opsonic (and presumably protective) antibodies in rabbits without evoking heart cross-reactive antibodies.

scholarly journals Location of variable and conserved epitopes among the multiple serotypes of streptococcal M protein.

1985 ◽  
Vol 161 (3) ◽  
pp. 623-628 ◽  
Author(s):  
K F Jones ◽  
B N Manjula ◽  
K H Johnston ◽  
S K Hollingshead ◽  
J R Scott ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Immune System ◽  
Cell Surface ◽  
Terminal Region ◽  
M Protein ◽  
Amino Terminal ◽  
Multiple Group ◽  
Streptococcal M Protein ◽  
Group A

In studies primarily confined to the amino-terminal region of the fibrillar group A streptococcal M protein, only limited immunological crossreactions have been observed among M serotypes. In this investigation, two monoclonal antibodies generated against nearly the entire M6 molecule (LysM6) were used to determine the extent of crossreactions among M serotyping strains and to localize their epitopes on the M molecule. Colony blot and immunoblot analyses revealed that an epitope responsible for crossreactions among 5 of the 56 strains of different M serotypes tested is located in the amino-terminal half of the molecule, distal to the cell surface. In contrast, a more common crossreactive epitope, reacting with 20 of the 56 strains, is located near the middle of the M molecule. These studies also reveal that the more conserved determinant, located more proximally to the cell surface, is accessible to the immune system, even on the whole organism, and, thus, may be useful in devising means to protect against infections by multiple group A streptococcal M serotypes.

scholarly journals Streptococcal M protein extracted by nonionic detergent. II. Analysis of the antibody response to the multiple antigenic determinants of the M-protein molecule.

1977 ◽  
Vol 146 (4) ◽  
pp. 1108-1123 ◽  
Author(s):  
V A Fischetti
Keyword(s):  
Solid Phase ◽  
Protein Molecule ◽  
The Other ◽  
M Protein ◽  
Antigenic Determinants ◽  
Specific Response ◽  
Human Beings ◽  
Inhibition Assay ◽  
Streptococcal M Protein ◽  
Nonionic Detergent

Purified streptococcal M protein extracted by nonionic detergent was used in an RIA and a solid-phase radiocompetitive inhibition assay to determine the nature of the immune response in both human beings and hyperimmunized rabbits to this complex antiphagocytic antigen. Results indicate that a type-specific response to an M antigen with the development of opsonic antibodies is the result of antibodies directed against the majority of the antigenic determinants of the molecule. Cross-reactions between certain M types on the other hand, are represented by antibodies directed against only a small percentage of these antigenic determinants. Results also suggest that avidity may play a role in the action of opsonic antibodies. However, the data indicate that factors besides avidity (i.e. sites bound by the antibodies) also seem essential for opsonization.

Peptide vaccine against canine parvovirus: identification of two neutralization subsites in the N terminus of VP2 and optimization of the amino acid sequence.

1995 ◽  
Vol 69 (11) ◽  
pp. 7274-7277 ◽  
Author(s):  
J I Casal ◽  
J P Langeveld ◽  
E Cortés ◽  
W W Schaaper ◽  
E van Dijk ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Peptide Vaccine ◽  
Canine Parvovirus ◽  
N Terminus

scholarly journals A Genetic Analysis of Interactions with Spc110p Reveals Distinct Functions of Spc97p and Spc98p, Components of the Yeast γ-Tubulin Complex

1998 ◽  
Vol 9 (8) ◽  
pp. 2201-2216 ◽  
Author(s):  
Thu Nguyen ◽  
Dani B.N. Vinh ◽  
Douglas K. Crawford ◽  
Trisha N. Davis
Keyword(s):  
Spindle Pole Body ◽  
Spindle Pole ◽  
Amino Terminus ◽  
Temperature Sensitive ◽  
Microtubule Organizing Center ◽  
Synthetic Lethal ◽  
Lethal Phenotype ◽  
Amino Terminal ◽  
N Terminus ◽  
Two Hybrid

The spindle pole body (SPB) in Saccharomyces cerevisiae functions as the microtubule-organizing center. Spc110p is an essential structural component of the SPB and spans between the central and inner plaques of this multilamellar organelle. The amino terminus of Spc110p faces the inner plaque, the substructure from which spindle microtubules radiate. We have undertaken a synthetic lethal screen to identify mutations that enhance the phenotype of the temperature-sensitive spc110–221 allele, which encodes mutations in the amino terminus. The screen identified mutations inSPC97 and SPC98, two genes encoding components of the Tub4p complex in yeast. The spc98–63allele is synthetic lethal only with spc110 alleles that encode mutations in the N terminus of Spc110p. In contrast, thespc97 alleles are synthetic lethal withspc110 alleles that encode mutations in either the N terminus or the C terminus. Using the two-hybrid assay, we show that the interactions of Spc110p with Spc97p and Spc98p are not equivalent. The N terminus of Spc110p displays a robust interaction with Spc98p in two different two-hybrid assays, while the interaction between Spc97p and Spc110p is not detectable in one strain and gives a weak signal in the other. Extra copies of SPC98 enhance the interaction between Spc97p and Spc110p, while extra copies of SPC97interfere with the interaction between Spc98p and Spc110p. By testing the interactions between mutant proteins, we show that the lethal phenotype in spc98–63 spc110–221 cells is caused by the failure of Spc98–63p to interact with Spc110–221p. In contrast, the lethal phenotype in spc97–62 spc110–221 cells can be attributed to a decreased interaction between Spc97–62p and Spc98p. Together, these studies provide evidence that Spc110p directly links the Tub4p complex to the SPB. Moreover, an interaction between Spc98p and the amino-terminal region of Spc110p is a critical component of the linkage, whereas the interaction between Spc97p and Spc110p is dependent on Spc98p.

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