PDBsum entry 2mf6

Biotin binding protein

PDB id: 2mf6

Contents

Protein chains

129 a.a.

PDB id:

2mf6

Name:

Biotin binding protein

Title:

Solution nmr structure of chimeric avidin, chiavd(i117y), in the biotin bound form

Structure:

Avidin, avidin-related protein 4/5. Chain: a, d, b, c. Fragment: p02701 residues 25-61, 85-152 and p56734 residues 62-82. Engineered: yes. Mutation: yes. Other_details: chimera of avidin and avidin-related protein 4/5

Source:

Gallus gallus. Chicken. Organism_taxid: 9031. Gene: avd, avr4, avr5. Expressed in: escherichia coli. Expression_system_taxid: 562.

NMR struc:

15 models

Authors:

H.Tossavainen,S.Kukkurainen,J.A.E.Maatta,T.Pihlajamaa,V.P.Hytonen, M.S.Kulomaa,P.Permi

Key ref:

H.Tossavainen et al. (2014). Chimeric Avidin--NMR structure and dynamics of a 56 kDa homotetrameric thermostable protein. Plos One, 9, e100564. PubMed id: 24959850 DOI: 10.1371/journal.pone.0100564

Date:

07-Oct-13 Release date: 06-Aug-14

Protein chains

P02701  (AVID_CHICK) -  Avidin from Gallus gallus

Seq: 152 a.a.

Struc: 129 a.a.*

Protein chains

P56734  (AVR4_CHICK) -  Avidin-related protein 4/5 from Gallus gallus

Seq: 150 a.a.

Struc: 129 a.a.*

* PDB and UniProt seqs differ at 34 residue positions (black crosses)

DOI no: 10.1371/journal.pone.0100564

Plos One 9:e100564 (2014)

PubMed id: 24959850

Chimeric Avidin--NMR structure and dynamics of a 56 kDa homotetrameric thermostable protein.

H.Tossavainen, S.Kukkurainen, J.A.Määttä, N.Kähkönen, T.Pihlajamaa, V.P.Hytönen, M.S.Kulomaa, P.Permi.

ABSTRACT

Chimeric avidin (ChiAVD) is a product of rational protein engineering remarkably resistant to heat and harsh conditions. In quest of the fundamentals behind factors affecting stability we have elucidated the solution NMR spectroscopic structure of the biotin-bound form of ChiAVD and characterized the protein dynamics through 15N relaxation and hydrogen/deuterium (H/D) exchange of this and the biotin-free form. To surmount the challenges arising from the very large size of the protein for NMR spectroscopy, we took advantage of its high thermostability. Conventional triple resonance experiments for fully protonated proteins combined with methyl-detection optimized experiments acquired at 58°C were adequate for the structure determination of this 56 kDa protein. The model-free parameters derived from the 15N relaxation data reveal a remarkably rigid protein at 58°C in both the biotin-bound and the free forms. The H/D exchange experiments indicate a notable increase in hydrogen protection upon biotin binding.