Structural aspects of thiol-specific spin labeling of human plasma low density lipoprotein.
A novel thiol-specific spin labeling procedure for the protein component (apoprotein B, apoB) of low density lipoproteins (LDLs) is presented. A methanethiosulfonate spin label was used to probe the free cysteine residues of apoB with electron paramagnetic resonance (EPR) spectroscopy. The results indicated that the spin labeled sites are predominantly buried in the LDL particle in two distinct environments that differ in their mobility restrictions. The suitability of thiol-specific labeling for the study of the stability and conformation of apoB was demonstrated in experiments with denaturing agents. The results presented in this work offer a new approach for the matching of EPR data with the primary structure of apoB.
Kveder
M
M
Ruder Bosković Institute, FK/LMR, Bijenicka 54, 10000 Zagreb, Croatia. kveder@rudjer.irb.hr
Pifat
G
G
Vukelić
B
B
Pecar
S
S
Schara
M
M
D001055:Apolipoproteins B / Q000097:blood / Q000737:chemistry; D003497:Cyclic N-Oxides; D004578:Electron Spin Resonance Spectroscopy; D006801:Humans; D008077:Lipoproteins, LDL / Q000097:blood / Q000737:chemistry*; D008698:Mesylates; D013113:Spin Labels; D013438:Sulfhydryl Compounds / Q000737:chemistry
D001055:Apolipoproteins B; D003497:Cyclic N-Oxides; D008077:Lipoproteins, LDL; D008698:Mesylates; D013113:Spin Labels; D013438:Sulfhydryl Compounds; C034248:(1-oxyl-2,2,5,5-tetramethylpyrroline-3-methyl)methanethiosulfonate