5KKM

Con-Vc11-22

Summary for 5KKM

• Entry DOI: 10.2210/pdb5kkm/pdb
• NMR Information: BMRB: 30124
• Descriptor: O2_contryphan_Vc1 prepropeptide (1 entity in total)
• Functional Keywords: single disulfide-directed beta hairpin, contryphan-vc1, stability, peptide scaffold, unknown function
• Biological source: Conus victoriae (Queen Victoria cone)
• Total number of polymer chains: 1
• Total formula weight: 2484.87

Authors

Chittoor, B.,Krishnarjuna, B.,MacRaild, C.A.,Robinson, S.D. (deposition date: 2016-06-22, release date: 2017-05-31, Last modification date: 2024-11-06)

Primary citation

Chittoor, B.,Krishnarjuna, B.,Morales, R.A.V.,MacRaild, C.A.,Sadek, M.,Leung, E.W.W.,Robinson, S.D.,Pennington, M.W.,Norton, R.S.
The Single Disulfide-Directed beta-Hairpin Fold. Dynamics, Stability, and Engineering.
Biochemistry, 56:2455-2466, 2017

PubMed Abstract: Grafting bioactive peptide sequences onto small cysteine-rich scaffolds is a promising strategy for enhancing their stability and value as novel peptide-based therapeutics. However, correctly folded disulfide-rich peptides can be challenging to produce by either recombinant or synthetic means. The single disulfide-directed β-hairpin (SDH) fold, first observed in contryphan-Vc1, provides a potential alternative to complex disulfide-rich scaffolds. We have undertaken recombinant production of full-length contryphan-Vc1 (rCon-Vc1[Z1Q]) and a truncated analogue (rCon-Vc1[Z1Q]), analyzed the backbone dynamics of rCon-Vc1[Z1Q], and probed the conformational and proteolytic stability of these peptides to evaluate the potential of contryphan-Vc1 as a molecular scaffold. Backbone N relaxation measurements for rCon-Vc1[Z1Q] indicate that the N-terminal domain of the peptide is ordered up to Thr19, whereas the remainder of the C-terminal region is highly flexible. The solution structure of truncated rCon-Vc1[Z1Q] was similar to that of the full-length peptide, indicating that the flexible C-terminus does not have any effect on the structured domain of the peptide. Contryphan-Vc1 exhibited excellent proteolytic stability against trypsin and chymotrypsin but was susceptible to pepsin digestion. We have investigated whether contryphan-Vc1 can accept a bioactive epitope while maintaining the structure of the peptide by introducing peptide sequences based on the DINNN motif of inducible nitric oxide synthase. We show that sCon-Vc1[NNN] binds to the iNOS-binding protein SPSB2 with an affinity of 1.3 μM while maintaining the SDH fold. This study serves as a starting point in utilizing the SDH fold as a peptide scaffold.

PubMed: 28437072
DOI: 10.1021/acs.biochem.7b00120

Experimental method

SOLUTION NMR