2MF6

Solution NMR structure of Chimeric Avidin, ChiAVD(I117Y), in the biotin bound form

Summary for 2MF6

• Entry DOI: 10.2210/pdb2mf6/pdb
• Related: 3MM0
• NMR Information: BMRB: 18125
• Descriptor: Avidin, Avidin-related protein 4/5 (1 entity in total)
• Functional Keywords: biotin binding protein
• Biological source: Gallus gallus (chicken)
• Cellular location: Secreted: P02701
• Total number of polymer chains: 4
• Total formula weight: 57601.02

Authors

Tossavainen, H.,Kukkurainen, S.,Maatta, J.A.E.,Pihlajamaa, T.,Hytonen, V.P.,Kulomaa, M.S.,Permi, P. (deposition date: 2013-10-07, release date: 2014-08-06, Last modification date: 2024-11-06)

Primary citation

Tossavainen, H.,Kukkurainen, S.,Maatta, J.A.E.,Kahkonen, N.,Pihlajamaa, T.,Hytonen, V.P.,Kulomaa, M.S.,Permi, P.
Chimeric Avidin - NMR Structure and Dynamics of a 56 kDa Homotetrameric Thermostable Protein
Plos One, 9:e100564-e100564, 2014

PubMed 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.

PubMed: 24959850
DOI: 10.1371/journal.pone.0100564

Experimental method

SOLUTION NMR