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PDBsum entry 2mf6
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Biotin binding protein
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PDB id
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2mf6
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References listed in PDB file
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Key reference
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Title
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Chimeric avidin--Nmr structure and dynamics of a 56 kda homotetrameric thermostable protein.
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Authors
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H.Tossavainen,
S.Kukkurainen,
J.A.Määttä,
N.Kähkönen,
T.Pihlajamaa,
V.P.Hytönen,
M.S.Kulomaa,
P.Permi.
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Ref.
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Plos One, 2014,
9,
e100564.
[DOI no: ]
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PubMed id
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Abstract
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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.
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