Peptide variants of protein A

Details Peptide variants of protein A Peptide variants of protein A

1998-03-06

2001-03-06

Woodward, Michael P. (Department: 1654)

Chemistry: natural resins or derivatives; peptides or proteins;

Peptides of 3 to 100 amino acid residues

Cyclic peptides

C530S324000, C530S350000, C514S011400, C514S012200, C424S192100, C424S193100, C424S194100

Reexamination Certificate

active

06197927

ABSTRACT:

FIELD OF THE INVENTION
This invention relates to the field of staphylococcal protein A, and more particularly to the gamma-immunoglobulin binding domains of protein A.
BACKGROUND OF THE INVENTION
Protein A from
Staphylococcus aureus
binds with high affinity and high specificity to the C&ggr;2-C&ggr;3 interface region of IgG (Langone,
Adv. Immunol
., 32: 157-252 (1982)). Protein A also exhibits an affinity for the Fab region of immunoglobulins that are encoded by the V
H
gene family, V
H
III (Sasso et al.,
J. Immunol,
61: 3026-3031 (1991); Hillson et al.,
A. J Exp. Med.,
178: 331-336 (1993)). The sequence of the gene coding for protein A revealed two functionally distinct regions (Uhlen et al.,
J. Biol. Chem.,
259: 1695-1702 (1984); Lofdahl et al.,
Proc. Natl. Acad Sci
(
USA
), 80: 697-701 (1983)). The amino-terminal region contains five highly homologous IgG-binding domains (termed E, D, A, B and C), and the carboxy terminal region anchors the protein to the cell wall and membrane. All five IgG-binding domains of protein A bind to IgG via the Fc region.
The structure of the B domain has been studied using
1
H-NMR (Torigoe et al.,
Biochem,
29: 8787-8793 (1990); Gouda et al.,
Biochem.,
31: 9665-9672 (1992)) and found to consist of three &agr;-helical regions (&agr;
1
, &agr;
2
, &agr;
3
corresponding to helices I, II and III in the NMR structure) which also are retained when bound to the Fc region of IgG (Gouda, supra). The tri-helical nature of the bound state is in contrast to the X ray crystal structure reported by Deisenhofer,
Biochem.,
20: 2361-2370 (1981) which showed that &agr;
1
and &agr;
2
helices of the B domain were present while the &agr;
3
helix did not form. The X ray crystallographic analysis reported by Deisenhofer, supra, and the mutagenesis studies reported by Popplewell et al.,
Protein Eng.,
4: 963-970 (1991) and Cedergren et al.,
Protein Eng.,
6: 441-448 (1993) have also identified ten residues within &agr;
1
and &agr;
2
and nine residues within the Fc region of IgG which participate in the protein-protein interaction.
The interaction between protein A and IgG forms the basis of many immunoaffinity-based purification procedures for antibodies (Hjelm,
FEBS Lett.,
28: 73-76 (1972)), antibody fragments and antigens (Sisson and Carter,
Immunol. Meth.,
127: 215-22- (1990)). Bottomley et al.,
J. Immunol. Meth.,
182: 185-192 (1995) reported that truncating B domain variant peptides lacking the 13 C-terminal residues of the &agr;
3
helix causes a decrease in IgG-binding activity. Bottomley et al. also characterized various mutations of residues within the &agr;
1
and &agr;
2
regions found to reduce or not affect IgG-binding activity.
SUMMARY OF THE INVENTION
In one aspect, the invention provides a compound represented by Formula (I):
X
1
-AA
6
-AA
7
-AA
8
-AA
9
-Gln-Gln-AA
12
-AA
13
-Phe-Tyr-Glu-Ala-Leu-His-Asp-Pro-Asn-Leu-Asn-Glu-Glu-Gln-Arg-Asn-Ala-Lys-Ile-AA
33
-Ser-Ile-AA
36
-Asp-Asp-X
2
(SEQ ID NO:1)  (I)
where
X
1
is selected from the group consisting of H, C
1
-C
6
alkanoyl, and Z-Ala-Val-AA
3
-AA
4
-AA
5
(SEQ ID NO:2);
where
Z is selected from the group consisting of H and C
1
-C
6
alkanoyl;
AA
3
is selected from the group consisting of Asp, Arg, and Ala;
AA
4
is selected from the group consisting of Asn and Gin; and
AA
5
is selected from the group consisting of Lys, Gly, and Ser;
AA
6
is selected from the group consisting of Phe and Gly;
AA
7
is selected from the group consisting of Asn and Trp;
AA
8
is selected from the group consisting of Lys and Met;
AA
9
is selected from the group consisting of Glu, Gln, and Arg;
AA
12
is selected from the group consisting of Asn, Ala, and Arg;
AA
13
is selected from the group consisting of Ala and Arg;
AA
33
is selected from the group consisting of Gin and Lys;
AA
36
is selected from the group consisting of Lys and Arg; and
X
2
is selected from the group consisting of OR
1
and NR
1
R
2
where R
1
and R
2
are independently selected from the group consisting of H, C
1
-C
6
alkyl, C
6
-C
12
aryl and C
6
-C
12
aryl-C
1
-C
6
alkyl.
In another aspect, the invention provides a compound represented by Formula (II):
where
X
1
is selected from the group consisting of H, C
1
-C
6
alkanoyl, and Z-Ala-Val-AA
3
-AA
4
-AA
5
(SEQ ID NO:2);
where
Z is selected from the group consisting of H and C
1
-C
6
alkanoyl;
AA
3
is selected from the group consisting of Asp, Arg, and Ala;
AA
4
is selected from the group consisting of Asn and Gln; and
AA
5
is selected from the group consisting of Lys, Gly, and Ser;
AA
6
is selected from the group consisting of Phe and Gly;
AA
7
is selected from the group consisting of Asn and Trp;
AA
8
is selected from the group consisting of Lys and Met;
AA
9
is selected from the group consisting of Glu, Gln, and Arg;
AA
12
is selected from the group consisting of Asn, Ala, and Arg;
AA
13
is selected from the group consisting of Ala and Arg;
AA
33
is selected from the group consisting of Gln and Lys;
AA
36
is selected from the group consisting of Lys and Arg; and
X
2
is selected from the group consisting of OR
1
and NR
1
R
2
where R
1
and R
2
are independently selected from the group consisting of H, C
1
-C
6
alkyl, C
6
-C
12
aryl and C
6
-C
12
aryl-C
1
-C
6
alkyl.


REFERENCES:
Rudinger, J ‘Characteristics of amino acids as components of peptide hormones sequence’ in Peptide Hormones, (ed. J.A. Parsons). University Park Press, Baltimore, pp. 1-7, 1976.
Braisted et al., ‘Minimizing a Binding Domain From Protein A’, Proc. Nat. Acad. Sci. 93:5688-5692, Jun. 1996.
Bottomley et al., “Elution of human IgG from affinity columns containing immobilised variants of protein A”Journal of Immunological Methods182:185-192 (1995).
Bottomley et al., “The stability and unfolding of an IgG binding protein based upon the B domain of protein A fromStaphylococcus aureusprobed by tryptophan substitution and fluorescence spectroscopy”Protein Eng. 7(12) :1463-1470 (1994).
Braisted & Wells, “Evolution of a Reduced Binding Epitope”96th General Meeting of the Amer. Society for Microbiology(Presentation summary) (May 22, 1996).
Braisted et al., “Minimizing a binding domain from protein A”Proc. Natl. Acad. Sci. 93:5688-5692 (Jun. 11, 1996).
Braisted, A., “Evolution of a Reduced Binding Epitope”IBC Conference of Applied Molecular Evolution(Presentation summary) (Dec. 7, 1995).
Cedergren et al., “Mutational analysis of the interaction between staphylococcal protein A and human IgG1”Protein Engineering6 (4) :441-448 (1993).
Diesenhofer, “Crystallographic Refinement and Atomic Models of a Human Fc Fragment and Its Complex with Fragment B of Protein A fromStaphylococcus aureusat 2.9-and 2.8-Angstroms Resolution”Biochemistry20 (9) :2361-2370 (1981).
Djojonegoro et al., “Bacteriophage Surface Display of an Immunoglobulin-binding Domain ofStaphylococcus aureusProtein A”Bio/Technology12:169-172 (1994).
Gouda et al., “Three-Dimensional Solution Structure of the B Domain of Staphylococcal Protein A: Comparisons of the Solution and Crystal Structures”Biochemistry31:9665-9672 (1992).
Hillson et al., “The Structural Basis of Germline-encoded VH3 Immunoglobulin-Binding to Staphylococcal Protein A”Journal of Experimental Medicine178:331-336 (1993).
Hjelm et al., “Protein A fromStaphylococcus aureus. Its isolation by affinity chromatography and its use as an immunosorbent for isolation of immunoglubulins”FEBS Letters28 (1):73-76 (1972).
Huston et al., “Multisite association by recombinant proteins can enhance binding selectivity”Biophysical Journal62:87-91 (1992).
Langone, “Protein A ofStaphylococcus aureusand Related Immunoglobulin Receptors Produced by Streptococci and Pneumonococci”Advances in Immunology32:157-252 (1982).
Lofdahl et al., “Gene for staphylococcal protein A”Proc. Natl. Acad. Sci.(USA) 80:697-701 (1983).
Nilsson et al., “A synthetic IgG-binding domain based on staphylococcal protein A”Protein Eng. 1:107-113 (1987).
Nord et al., “A combinational library of an alpha-helical bacterial receptor domain”Protein Engineering8 (6) :601-608 (1995).
Popplewell et al.,

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