Sequential Assignments and Identification of Secondary Structure Elements of the Colicin E9 Immunity Protein in Solution by Homonuclear and Heteronuclear NMR

Biochemistry ◽  
1994 ◽  
Vol 33 (41) ◽  
pp. 12347-12355 ◽  
Author(s):  
Michael J. Osborne ◽  
Lu-Yun Lian ◽  
Russell Wallis ◽  
Ann Reilly ◽  
Richard James ◽  
...  
Keyword(s):  
Secondary Structure ◽  
Heteronuclear Nmr ◽  
Immunity Protein ◽  
Colicin E9

scholarly journals In vivo and in vitro characterization of overproduced colicin E9 immunity protein

1992 ◽  
Vol 207 (2) ◽  
pp. 687-695 ◽  
Author(s):  
Russell WALLIS ◽  
Ann REILLY ◽  
Arthur ROWE ◽  
Geoffrey R. MOORE ◽  
Richard JAMES ◽  
...  
Keyword(s):  
Immunity Protein ◽  
In Vitro Characterization ◽  
Colicin E9

Dual Recognition and the Role of Specificity-Determining Residues in Colicin E9 DNase−Immunity Protein Interactions†

Biochemistry ◽  
1998 ◽  
Vol 37 (34) ◽  
pp. 11771-11779 ◽  
Author(s):  
Wei Li ◽  
Stefan J. Hamill ◽  
Andrew M. Hemmings ◽  
Geoffrey R. Moore ◽  
Richard James ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Immunity Protein ◽  
Colicin E9

scholarly journals Identification of critical residues in the colicin E9 DNase binding region of the Im9 protein

1997 ◽  
Vol 323 (3) ◽  
pp. 823-831 ◽  
Author(s):  
Michael J. OSBORNE ◽  
Russell WALLIS ◽  
Kit-Yi LEUNG ◽  
Glyn WILLIAMS ◽  
Lu-Yun LIAN ◽  
...  
Keyword(s):  
Binding Site ◽  
Protein Complexes ◽  
Dissociation Rate ◽  
15N Nmr ◽  
Binding Region ◽  
Nmr Studies ◽  
Immunity Protein ◽  
Wide Range ◽  
Protein Dissociation ◽  
Colicin E9

1H–15N NMR studies, in conjunction with mutagenesis experiments, have been used to delineate the DNase-binding surface of the colicin E9 inhibitor protein Im9 (where Im stands for immunity protein). Complexes were formed between the 15 kDa unlabelled E9 DNase domain and the 9.5 kDa Im9 protein uniformly labelled with 15N. Approx. 90% of the amide resonances of the bound Im9 were assigned and spectral parameters obtained from 1H–15N heteronuclear single quantum coherence (HSQC) spectra were compared with those for the free Im9 assigned previously. Many of the amide resonances were shifted on complex formation, some by more than 2 p.p.m. in the 15N dimension and more than 0.5 p.p.m. in the 1H dimension. Most of the strongly shifted amides are located on the surfaces of two of the four helices, helix II and helix III. Whereas helix II had already been identified through genetic and biochemical investigations as an important determinant of biological specificity, helix III had not previously been implicated in binding to the DNase. To test the robustness of the NMR-delineated DNase-binding site, a selection of Im9 alanine mutants were constructed and their dissociation rate constants from E9 DNase-immunity protein complexes quantified by radioactive subunit exchange kinetics. Their off-rates correlated well with the NMR perturbation analysis; for example, residues that were highly perturbed in HSQC experiments, such as residues 34 (helix II) and 54 (helix III), had a marked effect on the DNase–immunity protein dissociation rate when replaced by alanine. The NMR and mutagenesis data are consistent with a DNase-binding region on Im9 composed of invariant residues in helix III and variable residues in helix II. The relationship of this binding site model to the wide range of affinities (Kd values in the range 10-4 to 10-16 M) that have been measured for cognate and non-cognate colicin DNase–immunity protein interactions is discussed.

Specificity Determinants of the Interaction Between Colicin E9 and its Immunity Protein

1992 ◽  
pp. 181-201
Author(s):  
R. James ◽  
M. D. Curtis ◽  
R. Wallis ◽  
M. Osborne ◽  
C. Kleanthous ◽  
...  
Keyword(s):  
Immunity Protein ◽  
Colicin E9 ◽  
Specificity Determinants

scholarly journals Heteronuclear NMR assignments and secondary structure of the coiled coil trimerization domain from cartilage matrix protein in oxidized and reduced forms

1997 ◽  
Vol 6 (8) ◽  
pp. 1734-1745 ◽  
Author(s):  
Ronald Wiltscheck ◽  
Sonja A. Dames ◽  
Andrei T. Alexandrescu ◽  
Richard A. Kammerer ◽  
Therese Schulthess ◽  
...  
Keyword(s):  
Secondary Structure ◽  
Matrix Protein ◽  
Nmr Assignments ◽  
Heteronuclear Nmr ◽  
Coiled Coil ◽  
Cartilage Matrix ◽  
Heteronuclear Nmr Assignments ◽  
Reduced Forms

Protein-Protein Interactions in Colicin E9 DNase-Immunity Protein Complexes. 2. Cognate and Noncognate Interactions That Span the Millilmolar to Femptomolar Affinity Range

Biochemistry ◽  
1995 ◽  
Vol 34 (42) ◽  
pp. 13751-13759 ◽  
Author(s):  
Russell Wallis ◽  
Kit-Yi Leung ◽  
Ansgar J. Pommer ◽  
Hortense Videler ◽  
Geoffrey R. Moore ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Protein Complexes ◽  
Protein Protein Interactions ◽  
Immunity Protein ◽  
Colicin E9
Sign in / Sign up

Export Citation Format

Share Document