title: The endoproteinase furin contains two essential Ca2+ ions stabilizing its N-terminus and the unique S1 specificity pocket.
abstract: The mammalian prohormone/proprotein convertase (PC) furin is responsible for the maturation of a great variety of homeostatic but also many pathogenic proteins within the secretory pathway and the endosomal pathway and at the cell surface. Similar to other members of the PC family, furin requires calcium for catalytic activity. In a previous paper, the structural association of the catalytic and the P-domain of furin was shown and data were presented indicating two or three calcium-binding sites. The exact number and the three-dimensional localization of the essential calcium sites within furin have now been determined by collecting X-ray diffraction data on either side of the Ca K absorption edge and by calculating a novel type of double difference map from these anomalous scattering data. Two calcium ions were unambiguously identified: the purely structural Ca-1 also conserved in the bacterial digestive subtilisins and the Ca-2 site specific to PCs and essential for the formation of the P1 specificity-determining S1-binding pocket. In addition, these anomalous diffraction data show that no tightly bound K(+) sites exist in furin.
authors.affiliation: Max-Planck-Institut für Biochemie, Abteilung Strukturforschung, Am Klopferspitz 18A, 82152 Martinsried, Germany. than@biochem.mpg.de
authors.forename: Manuel E
authors.initials: ME
authors.lastname: Than
authors.forename: Stefan
authors.initials: S
authors.lastname: Henrich
authors.forename: Gleb P
authors.initials: GP
authors.lastname: Bourenkov
authors.forename: Hans D
authors.initials: HD
authors.lastname: Bartunik
authors.forename: Robert
authors.initials: R
authors.lastname: Huber
authors.forename: Wolfram
authors.initials: W
authors.lastname: Bode
mesh_terms: D002118:Calcium / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D003627:Data Interpretation, Statistical; D045683:Furin / Q000737:chemistry*; D008670:Metals / Q000737:chemistry; D011487:Protein Conformation; D013379:Substrate Specificity; D013381:Subtilisins / Q000737:chemistry
chemical_list: D008670:Metals; D013381:Subtilisins; D045683:Furin; D002118:Calcium