title: Ferrous Campylobacter jejuni truncated hemoglobin P displays an extremely high reactivity for cyanide - a comparative study.
abstract: Campylobacter jejuni hosts two hemoglobins (Hbs). The Camplylobacter jejuni single-domain Hb (called Cgb) is homologous to the globin domain of flavohemoglobin, and it has been proposed to protect the bacterium against nitrosative stress. The second Hb is called Ctb (hereafter Cj-trHbP), belongs to truncated Hb group III, and has been hypothesized to be involved in O(2) chemistry. Here, the kinetics and thermodynamics of cyanide binding to ferric and ferrous Cj-trHbP [Cj-trHbP(III) and Cj-trHbP(II), respectively] are reported and analyzed in parallel with those of related heme proteins, with particular reference to those from Mycobacterium tuberculosis. The affinity of cyanide for Cj-trHbP(II) is higher than that reported for any known (in)vertebrate globin by more than three orders of magnitude (K = 1.2 x 10(-6) m). This can be fully attributed to the highest (ever observed for a ferrous Hb) cyanide-binding association rate constant (k(on) = 3.3 x 10(3) m(-1).s(-1)), even though the binding process displays a rate-limiting step (k(max) = 9.1 s(-1)). Cj-trHbP(III) shows a very high affinity for cyanide (L = 5.8 x 10(-9) m); however, cyanide association kinetics are independent of cyanide concentration, displaying a rate-limiting step (l(max) = 2.0 x 10(-3) s(-1)). Values of the first-order rate constant for cyanide dissociation from Cj-trHbP(II)-cyanide and Cj-trHbP(III)-cyanide (k(off) =5.0 x 10(-3) s(-1) and l(off) > or = 1 x 10(-4) s(-1), respectively) are similar to those reported for (in)vertebrate globins. The very high affinity of cyanide for Cj-trHbP(II), reminiscent of that of horseradish peroxidase(II), suggests that this globin may participate in cyanide detoxification.
authors.affiliation: Dipartimento di Biologia, Università Roma Tre, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Bolli
authors.forename: Chiara
authors.initials: C
authors.lastname: Ciaccio
authors.forename: Massimo
authors.initials: M
authors.lastname: Coletta
authors.forename: Marco
authors.initials: M
authors.lastname: Nardini
authors.forename: Martino
authors.initials: M
authors.lastname: Bolognesi
authors.forename: Alessandra
authors.initials: A
authors.lastname: Pesce
authors.forename: Michel
authors.initials: M
authors.lastname: Guertin
authors.forename: Paolo
authors.initials: P
authors.lastname: Visca
authors.forename: Paolo
authors.initials: P
authors.lastname: Ascenzi
mesh_terms: D000465:Algorithms; D016123:Campylobacter jejuni / Q000378:metabolism*; D003486:Cyanides / Q000378:metabolism*; D006418:Heme / Q000378:metabolism; D006454:Hemoglobins / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013816:Thermodynamics; D054793:Truncated Hemoglobins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D003486:Cyanides; D006454:Hemoglobins; D054793:Truncated Hemoglobins; C036602:hemoglobin P; D006418:Heme