title: Coupled responses of the regions near cysteine-190 and the carboxy terminus of rabbit cardiac tropomyosin: fluorescence and circular dichroism studies.
abstract: Rabbit cardiac tropomyosin was labeled at Cys-190 with either N-(1-pyrenyl)iodoacetamide (Py) or 6-acryloyl-2-(dimethylamino)naphthalene (AD, acrylodan). Half of the labeled sample then was treated with carboxypeptidase A to produce an identically labeled nonpolymerizable form of tropomyosin, NPTM. Investigation of temperature-dependent changes in pyrene excimer emission, acrylodan fluorescence polarization, and tyrosyl circular dichroism in different samples of tropomyosin and NPTM reveals that absence of the COOH-terminal portion of tropomyosin modifies the response of the Cys-190 region to heat. Removal of the COOH terminus releases certain conformational constraints from the coiled-coil back to and including the Cys-190 region without causing a severe drop in the net alpha-helical content of the protein. Observation of changes in pyrene excimer fluorescence and in fluorescence polarization of acrylodan with time after addition of carboxypeptidase A to samples of labeled tropomyosin directly demonstrates this relaxation process. Thermally induced reduction in tyrosyl circular dichroism, together with consideration of the distribution of tyrosyl residues on tropomyosin, also supports the proposal.
authors: Department of Chemistry, University of British Columbia, Vancouver, Canada.
authors: I D
authors: ID
authors: Clark
authors: L D
authors: LD
authors: Burtnick
mesh_terms: D000818:Animals; D002268:Carboxypeptidases / Q000378:metabolism / Q000494:pharmacology; D043422:Carboxypeptidases A; D002942:Circular Dichroism; D003545:Cysteine; D005454:Fluorescence Polarization; D009206:Myocardium / Q000737:chemistry*; D009994:Osmolar Concentration; D011487:Protein Conformation; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D014335:Tropomyosin / Q000737:chemistry* / Q000378:metabolism; D014443:Tyrosine
chemical_list: D014335:Tropomyosin; D014443:Tyrosine; D002268:Carboxypeptidases; D043422:Carboxypeptidases A; D003545:Cysteine