Magnetic Resonance Studies of Apolipoprotein C-I Nitroxide Labeled or [13C]Methyl Enriched at Methionine-38

Tsung Chang Chen, Roger D. Knapp, Michael F. Rohde, James R. Brainard, Antonio Gotto, James T. Sparrow, Joel D. Morrisett

Research output: Contribution to journalArticlepeer-review

12 Scopus citations


One of the three proposed lipid-binding regions of the human apolipoprotein C-I (apoC-I) is an amphipathic helix which extends from residue 33 to residue 53 and includes a single methionine at sequence position 38. The involvement of the sequence around methionine-38 in phospholipid binding has been evaluated with paramagnetic and nuclear reporter groups attached to the thiomethyl moiety. This moiety has been spin-labeled with N-(2,2,6,6-tetramethylpiperidinyl-1-oxy)bromoacetamide or 13C enriched with 13CH3I. As determined from its EPR spectrum, the nitroxide at Met-38 of apoC-I had a rotational correlation time (τc) of 0.22 ns. When dimyristoylphosphatidylcholine (DMPC) was bound to the spin-labeled apoprotein, τc increased to 0.35 ns, indicating decreased motion for the methionyl side chain. The line width (ν1/2) and spin-lattice relaxation time (T1) for the thiomethyl resonance of 13C-enriched apoC-I in 10 mM phosphate buffer was 6.0 Hz and 320 ms, respectively. When the protein solution was made 1.6 M in Gdn·HCl, these values changed to 2.6 Hz and 970 ms, respectively. Upon addition of DMPC multilamellar liposomes to [13C]apoC-I in 1.6 M Gdn·HCl, the line width increased to 4.7 Hz and the T1 decreased to 380 ms. These results strongly suggest that methionine-38 of apoC-I resides in a region of the apoprotein which undergoes significant secondary and/or tertiary structural change upon disaggregation/unfolding in Gdn·HCl and upon interaction with phospholipid.

Original languageEnglish (US)
Pages (from-to)5140-5146
Number of pages7
Issue number22
StatePublished - Feb 1 1980

ASJC Scopus subject areas

  • Biochemistry


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