title: The sequence location of the actin metal.
abstract: We have incorporated Fe2+ into the high-affinity metal-ion-binding site of actin. By supplying the system with oxygen from air and a reductant (dithiothreitol or ascorbate), we have induced free-radical generation, with the intent of causing peptide cleavage at the metal-ion-binding site. By analysis of the resulting fragments from actin in the F-form, we have deduced that cuts occurred at positions 159-160 and 301-302 (at the latter location we could not be sure if more than one cut occurred). We considered that these two cuts occurred in the chain strand coursing from the outer to the inner domain and vice-versa. Our results harmonize very well with the recently reported atomic structure of actin [Kabsch, W., Mannherz, H.G., Suck, D., Pai, E.F. & Holmes, K.C. (1990) Nature 347, 37-44] and remove ambiguities that had remained in the structure. The results partly bear out the homology-based prediction of Strzelecka-Golaszewska et al. [Strzelecka-Golaszewska, H., Boguta, G., Zmorzynshi, S. & Moraczcwska, J. (1989) Eur. J. Biochem. 182, 299-305].
authors.affiliation: Department of Physiology and Biophysics, University of the Pacific, San Francisco 94115.
authors.forename: K
authors.initials: K
authors.lastname: Ue
authors.forename: A
authors.initials: A
authors.lastname: Muhlrad
authors.forename: C G
authors.initials: CG
authors.lastname: Edmonds
authors.forename: D
authors.initials: D
authors.lastname: Bivin
authors.forename: A
authors.initials: A
authors.lastname: Clark
authors.forename: W V
authors.initials: WV
authors.lastname: Piechowski
authors.forename: M F
authors.initials: MF
authors.lastname: Morales
mesh_terms: D000199:Actins / Q000302:isolation & purification / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D002413:Cations, Divalent; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D007501:Iron / Q000378:metabolism; D013058:Mass Spectrometry; D008670:Metals / Q000378:metabolism*; D010084:Oxidation-Reduction; D010455:Peptides / Q000378:metabolism; D011817:Rabbits
chemical_list: D000199:Actins; D002413:Cations, Divalent; D008670:Metals; D010455:Peptides; D007501:Iron; D000251:Adenosine Triphosphatases; D002118:Calcium