Species-specific differences in covalently crosslinked complexes of yeast cytochrome c peroxidase with horse and yeast iso-1 ferricytochromes c.
1. The results of chemically crosslinking yeast cytochrome c peroxidase with both horse and yeast iso-1 ferricytochromes c have been studied by a combination of gel electrophoresis and proton NMR spectroscopy. 2. The complexes were formed at a variety of potassium phosphate concentrations ranging from 10 to 300 mM using the water soluble crosslinking agent, EDC (1-ethyl-3-[3-(dimethylamino)propyl]-carbodiimide). 3. The primary crosslinking product in both cases is the 1:1 covalent complex, but, for each pair of partner proteins the yield of the 1:1 crosslinked complex varies with the salt concentration. 4. Furthermore, at low salt concentrations the yield of the 1:1 covalent complex involving horse cytochrome c is much larger than the yield of the 1:1 covalent complex formed with yeast iso-1 cytochrome c, whereas at high salt concentrations the situation is reversed. 5. Proton NMR spectroscopy, in combination with gel electrophoresis, provides evidence for the formation of different types of 1:1 complexes for the peroxidase/yeast cytochrome c pair and has been used to study the effect of changes in the solution ionic strength upon both the peroxidases/horse cytochrome c and the peroxidase/yeast cytochrome c complexes. 6. This work indicates that electrostatic interactions between proteins play a dominant role in formation of complexes between cytochrome c peroxidase and horse ferricytochrome c, whereas the hydrophobic effect plays a comparatively larger role in stabilizing complexes between cytochrome c peroxidase and yeast iso-1 ferricytochrome c.
Moench
S J
SJ
Department of Chemistry, Northern Illinois University, DeKalb 60115.
Erman
J E
JE
Satterlee
J D
JD
D000818:Animals; D003432:Cross-Linking Reagents; D003574:Cytochrome c Group / Q000737:chemistry*; D003578:Cytochrome-c Peroxidase / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D005022:Ethyldimethylaminopropyl Carbodiimide; D006736:Horses / Q000378:metabolism*; D007527:Isoenzymes / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D009994:Osmolar Concentration; D010649:Phenylalanine / Q000737:chemistry; D011522:Protons; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012996:Solutions; D013045:Species Specificity
D003432:Cross-Linking Reagents; D003574:Cytochrome c Group; D007527:Isoenzymes; D011522:Protons; D012996:Solutions; D010649:Phenylalanine; D003578:Cytochrome-c Peroxidase; D005022:Ethyldimethylaminopropyl Carbodiimide