Related ArticlesSolution secondary structure of a bacterially expressed peptide from the receptor binding domain of Pseudomonas aeruginosa pili strain PAK: A heteronuclear multidimensional NMR study.
The C-terminal receptor binding region of Pseudomonas aeruginosa pilin protein strain PAK (residues 128-144) has recently been the target for the design of a synthetic peptide vaccine effective against multiple strains of P. aeruginosa infection. We have successfully cloned and bacterially expressed a 15N-labeled PAK pilin peptide spanning residues 128-144 of the intact PAK pilin protein, PAK 128-144(Hs145), and have determined the solution secondary structure of this peptide using heteronuclear multidimensional NMR spectroscopy. The oxidized recombinant peptide exists as a major (trans) and minor (cis) species in solution, arising from isomerization around the Ile138-Pro139 peptide bond. The pattern of NOEs, temperature coefficients, and coupling constants observed for the trans isomer demonstrate the presence of a type I beta-turn and a type II beta-turn spanning Asp134-Glu-Gln-Phe137 and Pro139-Lys-Gly-Cys142, respectively. This is in agreement with the NMR solution structure of the trans isomer of a synthetic PAK 128-144 peptide which showed a type I and a type II beta-turn in these same regions of the sequence [McInnes, C., Sönnichsen, F. D., Kay, C. M., Hodges, R. S., and Sykes, B. D. (1993) Biochemistry 32, 13432-13440; Campbell, A. P., McInnes, C., Hodges, R. S., and Sykes, B. D. (1995) Biochemistry 34, 16255-16268]. The pattern of NOEs, temperature coefficients, and coupling constants observed for the cis isomer also demonstrate a type II beta-turn spanning Pro139-Lys-Gly-Cys142, but suggest a second beta-turn spanning Asp132-Gln-Asp-Glu135. Thus, the cis isomer may also possess a double-turn motif (like the trans isomer), but with different spacing between the turns and a different placement of the first turn in the sequence. The discovery of a double-turn motif in the trans (and cis) recombinant PAK pilin peptide is an extremely important result since the double turn has been implicated as a structural requirement for the recognition of both receptor and antibody. These results pave the way for future isotope-edited NMR studies of the labeled recombinant PAK pilin peptide bound to antibody and receptor, studies integral to the design of an effective synthetic peptide vaccine.
[NMR paper] NMR screening and crystal quality of bacterially expressed prokaryotic and eukaryotic
NMR screening and crystal quality of bacterially expressed prokaryotic and eukaryotic proteins in a structural genomics pipeline.
Related Articles NMR screening and crystal quality of bacterially expressed prokaryotic and eukaryotic proteins in a structural genomics pipeline.
Proc Natl Acad Sci U S A. 2005 Feb 8;102(6):1901-5
Authors: Page R, Peti W, Wilson IA, Stevens RC, Wüthrich K
In the Joint Center for Structural Genomics, one-dimensional (1D) 1H NMR spectroscopy is routinely used to characterize the folded state of protein targets and,...
nmrlearner
Journal club
0
11-24-2010 11:14 PM
[NMR paper] NMR structure and dynamics of a receptor-active apolipoprotein E peptide.
NMR structure and dynamics of a receptor-active apolipoprotein E peptide.
Related Articles NMR structure and dynamics of a receptor-active apolipoprotein E peptide.
J Biol Chem. 2002 Aug 9;277(32):29172-80
Authors: Raussens V, Slupsky CM, Ryan RO, Sykes BD
Apolipoprotein E (apoE) is important in lipid metabolism due to its interaction with members of the low density lipoprotein (LDL) receptor family. ApoE is able to interact with the LDL receptor only when it is bound to lipid particles. To address structural aspects of this phenomenon, a...
nmrlearner
Journal club
0
11-24-2010 08:49 PM
[NMR paper] Backbone dynamics of a bacterially expressed peptide from the receptor binding domain
Backbone dynamics of a bacterially expressed peptide from the receptor binding domain of Pseudomonas aeruginosa pilin strain PAK from heteronuclear 1H-15N NMR spectroscopy.
Related Articles Backbone dynamics of a bacterially expressed peptide from the receptor binding domain of Pseudomonas aeruginosa pilin strain PAK from heteronuclear 1H-15N NMR spectroscopy.
J Biomol NMR. 2000 Jul;17(3):239-55
Authors: Campbell AP, Spyracopoulos L, Irvin RT, Sykes BD
The backbone dynamics of a 15N-labeled recombinant PAK pilin peptide spanning residues...
nmrlearner
Journal club
0
11-19-2010 08:29 PM
[NMR paper] NMR solution structure of the receptor binding domain of human alpha(2)-macroglobulin
NMR solution structure of the receptor binding domain of human alpha(2)-macroglobulin.
Related Articles NMR solution structure of the receptor binding domain of human alpha(2)-macroglobulin.
J Biol Chem. 2000 Jan 14;275(2):1089-94
Authors: Huang W, Dolmer K, Liao X, Gettins PG
Human alpha(2)-macroglobulin-proteinase complexes bind to their receptor, the low density lipoprotein receptor-related protein (LRP), through a discrete 138-residue C-terminal receptor binding domain (RBD), which also binds to the beta-amyloid peptide. We have used NMR...
nmrlearner
Journal club
0
11-18-2010 09:15 PM
[NMR paper] Studies on solution NMR structure of brazzein : Secondary structure and molecular sca
Studies on solution NMR structure of brazzein : Secondary structure and molecular scaffold.
Related Articles Studies on solution NMR structure of brazzein : Secondary structure and molecular scaffold.
Sci China C Life Sci. 1999 Aug;42(4):409-19
Authors: Gao G, Dai J, Ding M, Hellekant G, Wang J, Wang D
Brazzein is a sweet-tasting protein isolated from the fruit of West African plantPentadiplandra brazzeana Baillon. It is the smallest and the most water-soluble sweet protein discovered so far and is highly thermostable. The proton NMR study of...
nmrlearner
Journal club
0
11-18-2010 08:31 PM
[NMR paper] Proton NMR sequence-specific assignments and secondary structure of a receptor bindin
Proton NMR sequence-specific assignments and secondary structure of a receptor binding domain of mouse gamma-interferon.
Related Articles Proton NMR sequence-specific assignments and secondary structure of a receptor binding domain of mouse gamma-interferon.
Biochemistry. 1993 Jun 1;32(21):5650-5
Authors: Sakai TT, Jablonsky MJ, DeMuth PA, Krishna NR, Jarpe MA, Johnson HM
Previous studies using synthetic peptides and monoclonal antibodies have implicated the N-terminal 39-residue segment as a receptor binding region of mouse gamma-interferon...
nmrlearner
Journal club
0
08-21-2010 11:53 PM
[NMR paper] NMR structure of a receptor-bound G-protein peptide.
NMR structure of a receptor-bound G-protein peptide.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.nature.com-images-lo_nature.gif Related Articles NMR structure of a receptor-bound G-protein peptide.
Nature. 1993 May 20;363(6426):276-81
Authors: Dratz EA, Furstenau JE, Lambert CG, Thireault DL, Rarick H, Schepers T, Pakhlevaniants S, Hamm HE
Heterotrimeric GTP-binding proteins (G proteins) regulate cellular activity by coupling to hormone or sensory receptors. Stimulated receptors catalyse the release of GDP from G protein...
nmrlearner
Journal club
0
08-21-2010 11:53 PM
[NMR paper] Secondary structure and topology of interleukin-1 receptor antagonist protein determi
Secondary structure and topology of interleukin-1 receptor antagonist protein determined by heteronuclear three-dimensional NMR spectroscopy.
Related Articles Secondary structure and topology of interleukin-1 receptor antagonist protein determined by heteronuclear three-dimensional NMR spectroscopy.
Biochemistry. 1992 Jun 16;31(23):5237-45
Authors: Stockman BJ, Scahill TA, Roy M, Ulrich EL, Strakalaitis NA, Brunner DP, Yem AW, Deibel MR
Interleukin-1 (IL-1) proteins, such as IL-1 beta, play a key role in immune and inflammatory responses....