Acid-induced conformational changes in calcium-depleted horseradish peroxidase

INOR 194

Jesse A. Pfammatter, jpfamm@gmail.com and Scott M. Tremain. Department of Chemistry, Eastern Illinois University, 600 Lincoln Ave., Charleston, IL 61920
The crystal structure of horseradish peroxidase (HRP) reveals two bound Ca2+ ions possibly required for catalysis and protein stability. We investigate acid-induced conformational changes in Ca2+-bound and Ca2+-depleted HRP by monitoring heme Soret absorbance. Soret band spectral changes at acidic pH over time depend on the anion present in solution. Upon dissolving Ca2+-bound HRP in water adjusted to pH 3 with HCl, the Soret band remained unchanged. In 50 mM phosphate buffer adjusted to pH 3 with HCl, the Soret band rapidly shifted from 403 nm to 370 nm. Phosphate amplifies acid-induced conformational changes in HRP. To investigate the role of Ca2+ in acid-induced conformational changes, Ca2+-depleted HRP was prepared. Surprisingly, acid-induced conformational changes in Ca2+-depleted HRP were similar to those in Ca2+-bound HRP. Acid-induced conformational changes in HRP appear to be more dependent on the anion than on the presence or absence of bound Ca2+.
 

The Bioinorganic Chemistry Relating to Enzymes and Proteins
7:00 PM-10:00 PM, Sunday, March 25, 2007 Hyatt Regency Chicago -- Riverside Center, Poster

Sci-Mix
8:00 PM-10:00 PM, Monday, March 26, 2007 Hyatt Regency Chicago -- Riverside Center, Sci-Mix

Division of Inorganic Chemistry

The 233rd ACS National Meeting, Chicago, IL, March 25-29, 2007