Structure of membrane-embedded M13 major coat protein is insensitive to hydrophobic stress.
The structure of a membrane-embedded alpha-helical reference protein, the M13 major coat protein, is characterized under different conditions of hydrophobic mismatch using fluorescence resonance energy transfer in combination with high-throughput mutagenesis. We show that the structure is similar in both thin (14:1) and thick (20:1) phospholipid bilayers, indicating that the protein does not undergo large structural rearrangements in response to conditions of hydrophobic mismatch. We introduce a "helical fingerprint" analysis, showing that amino acid residues 1-9 are unstructured in both phospholipid bilayers. Our findings indicate the presence of pi-helical domains in the transmembrane segment of the protein; however, no evidence is found for a structural adaptation to the degree of hydrophobic mismatch. In light of current literature, and based on our data, we conclude that aggregation (at high protein concentration) and adjustment of the tilt angle and the lipid structure are the dominant responses to conditions of hydrophobic mismatch.
Vos
Werner L
WL
Laboratory of Biophysics, Wageningen University, Wageningen, The Netherlands.
Schor
Marieke
M
Nazarov
Petr V
PV
Koehorst
Rob B M
RB
Spruijt
Ruud B
RB
Hemminga
Marcus A
MA
D000596:Amino Acids / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism; D003545:Cysteine / Q000737:chemistry; D008055:Lipids / Q000737:chemistry; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D015233:Models, Statistical; D008968:Molecular Conformation; D016296:Mutagenesis; D009154:Mutation; D009219:Myosin-Light-Chain Kinase / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry*; D010743:Phospholipids / Q000737:chemistry; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary
D000596:Amino Acids; D008055:Lipids; C107660:M13 protein (myosin light-chain kinase); D010446:Peptide Fragments; D010743:Phospholipids; D009219:Myosin-Light-Chain Kinase; D003545:Cysteine
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