炎症反应在缺氧中作用.pdfVIP

5266 Biochemistry 2008, 47, 5266–5270 Zebrafish Neuroglobin Is a Cell-Membrane-Penetrating Globin† Seiji Watanabe‡ and Keisuke Wakasugi*,‡,§ Department of Life Sciences, Graduate School of Arts and Sciences, The Uni Versity of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan, and Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology (JST), 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan Recei Ved February 18, 2008; ReVised Manuscript ReceiVed March 26, 2008 ABSTRACT : Neuroglobin (Ngb) is a recently discovered vertebrate heme protein that is expressed in the brain and can reversibly bind oxygen. Mammalian Ngb is involved in neuroprotection under oxidative stress conditions, such as ischemia and reperfusion. We previously demonstrated that human ferric Ngb binds to the R subunit of heterotrimeric G proteins (GRi) and acts as a guanine nucleotide dissociation inhibitor (GDI) for GRi . Recently, we used a protein delivery reagent, Chariot, and demonstrated that the GDI activity of human Ngb is tightly correlated with its neuroprotective activity. In the present study, we found that chimeric ZHHH Ngb, in which module M1 of human Ngb is replaced by that of zebrafish Ngb, protects PC12 cells against oxidative stress-induced cell death even in the absence of Chariot. Using fluorescein isothiocyanate (FITC)-labeled Ngb proteins, we demonstrated that both zebrafish and chimeric ZHHH Ngb can penetrate cell membranes in the absence of Chariot, suggesting that module M1 of zebrafish Ngb can translocate into cells. This is the first report of a native cell-membrane-penetrating globin. Neuroglobin (Ngb)1 is a heme protein, recent