Three amino acids that are critical to formation and stability of the P22 tailspike trimer.


Abstract

The P22 tailspike protein folds by forming a folding competent monomer species that forms a dimeric, then a non-native trimeric (protrimer) species by addition of folding competent monomers. We have found three residues, R549, R563, and D572, which play a critical role in both the stability of the native tailspike protein and assembly and maturation of the protrimer. King and colleagues reported previously that substitution of R563 to glutamine inhibited protrimer formation. We now show that the R549Q and R563K variants significantly delay the protrimer-to-trimer transition both in vivo and in vitro. Previously, variants that destabilize intermediates have shown wild-type chemical stability. Interestingly, both the R549Q and R563K variants destabilize the tailspike trimer in guanidine denaturation studies, indicating that they represent a new class of tailspike folding variants. R549Q has a midpoint of unfolding at 3.2M guanidine, compared to 5.6M for the wild-type tailspike protein, while R563K has a midpoint of unfolding of 1.8 M. R549Q and R563K also denature over a broader pH range than the wild-type tailspike protein and both proteins have increased sensitivity to pH during refolding, suggesting that both residues are involved in ionic interactions. Our model is that R563 and D572 interact to stabilize the adjacent turn, aiding the assembly of the dimer and protrimer species. We believe that the interaction between R563 and D572 is also critical following assembly of the protrimer to properly orient D572 in order to form a salt bridge with R549 during protrimer maturation. Study holds ProTherm entries: 19610, 19611, 19612 Extra Details: P22 tailspike; protein folding; stability; mutation; ionic interaction

Submission Details

ID: oTyKCJxM

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:51 p.m.

Version: 1

Publication Details
Gage MJ;Zak JL;Robinson AS,Protein Sci. (2005) Three amino acids that are critical to formation and stability of the P22 tailspike trimer. PMID:16081648
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
2VFO 2008-12-16 1.5 Low Temperature Structure of P22 Tailspike Protein Fragment (109-666), Mutant V125L
2VFM 2008-12-16 1.5 Low Temperature Structure of P22 Tailspike Protein Fragment (109-666)
2VFN 2008-12-16 1.5 Low Temperature Structure of P22 Tailspike Protein Fragment (109-666), Mutant V125A
2VFQ 2008-12-16 1.55 Low Temperature Structure of P22 Tailspike Protein Fragment (109-666), Mutant V450A
2VFP 2008-12-16 1.55 Low Temperature Structure of P22 Tailspike Protein Fragment (109-666), Mutant V349L
2XC1 2011-05-04 1.65 Full-length Tailspike Protein Mutant Y108W of Bacteriophage P22
3TH0 2012-08-29 1.75 P22 Tailspike complexed with S.Paratyphi O antigen octasaccharide
1TYX 1997-07-23 1.8 TITLE OF TAILSPIKE-PROTEIN
1TYU 1997-07-23 1.8 STRUCTURE OF TAILSPIKE-PROTEIN
1QQ1 2000-04-12 1.8 TAILSPIKE PROTEIN, MUTANT E359G
2VNL 2009-02-10 1.8 MUTANT Y108Wdel OF THE HEADBINDING DOMAIN OF PHAGE P22 TAILSPIKE C- TERMINally fused to ISOLEUCINE ZIPPER pIIGCN4 (chimera II)
1TYW 1997-07-23 1.8 STRUCTURE OF TAILSPIKE-PROTEIN
1TYV 1997-07-23 1.8 STRUCTURE OF TAILSPIKE-PROTEIN
5GAI 2016-02-17 10.5 Probabilistic Structural Models of Mature P22 Bacteriophage Portal, Hub, and Tailspike proteins
1TSP 1995-09-15 2.0 CRYSTAL STRUCTURE OF P22 TAILSPIKE PROTEIN: INTERDIGITATED SUBUNITS IN A THERMOSTABLE TRIMER
1CLW 1999-11-05 2.0 TAILSPIKE PROTEIN FROM PHAGE P22, V331A MUTANT
1QRB 2000-04-12 2.0 PLASTICITY AND STERIC STRAIN IN A PARALLEL BETA-HELIX: RATIONAL MUTATIONS IN P22 TAILSPIKE PROTEIN
1QA3 2000-01-12 2.0 TAILSPIKE PROTEIN, MUTANT A334I
1QA1 2000-01-12 2.0 TAILSPIKE PROTEIN, MUTANT V331G
1QA2 2000-01-12 2.0 TAILSPIKE PROTEIN, MUTANT A334V
2VKY 2009-02-10 2.05 Headbinding Domain of Phage P22 Tailspike C-Terminally Fused to Isoleucine Zipper pIIGCN4 (Chimera I)
1QRC 2000-04-12 2.5 TAILSPIKE PROTEIN, MUTANT W391A
1LKT 1998-01-28 2.6 CRYSTAL STRUCTURE OF THE HEAD-BINDING DOMAIN OF PHAGE P22 TAILSPIKE PROTEIN

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Tail spike protein P12528 FIBER_BPP22