caa a22 4500 477136257 CHVBK 20180405111718.0 cr unu---uuuuu 170330e19970101xx s 000 0 eng 10.1023/A:1018619501038 doi (NATIONALLICENCE)springer-10.1023/A:1018619501038 Resonance assignment and secondary structure determination and stability of the recombinant human uteroglobin with heteronuclear multidimensional NMR [Elektronische Daten] [Teresa Carlomagno, Giuditta Mantile, Renzo Bazzo, Lucio Miele, Livio Paolillo, Anil Mukherjee, Gaetano Barbato] Human uteroglobin (h-UG) or Clara cell 10kDa (cc10kDa) is a steroid-dependent, 17 kDahomodimeric, secretory protein with potent anti-inflammatory/immunomodulatory properties.However, the exact physiological role still remains to be determined. It has been hypothesisedthat its activity is exerted through the binding of a specific target represented by a smallmolecule (still unknown), and that the binding is regulated by the formation/disruption of twocysteine bonds. The binding properties of the reduced UG have been proved in vitro forseveral different molecules, but no in vivo data are available to date. However, binding hasbeen observed between reduced rabbit UG and a protein of an apparent molecular mass of90 kDa and, more recently, we found an h-UG-binding protein (putative receptor), of anapparent molecular mass of 190 kDa, on the surface of several cell types. The recognitioninvolves oxidised h-UG. These findings pose the problem of the relevance of the oxidationstate in the recognition process. To determine the solution structure of the oxidised h-UG, weproduced wild-type as well as uniformly 15N- and 15N/13C-labelled samples of therecombinant protein. The assignments of the 1H, 15N and 13C resonances are presented,based on a series of homonuclear 2D and 3D and heteronuclear 2D and 3D double and tripleresonance NMR experiments. Our results indicate that h-UG is an extremely stable proteinunder a wide range of temperatures and pH conditions. The secondary structure in solutionis in general agreement with previously reported crystal structures of rabbit UG, suggestingthat cc10kDa and h-UG are indeed the same protein. Small local differences found in the N-and C-terminal helices seem to support the hypothesis that flexibility involves these residues;moreover, it possibly accounts for the residual binding properties observed when the proteinis in the oxidised state. Kluwer Academic Publishers, 1997 Secondary structure nationallicence Uteroglobin nationallicence h-cc10kDa nationallicence Carlomagno Teresa Department of Chemistry, University of Naples 'Federico II‘, Via Mezzocannone 4, I-80100, Naples, Italy aut Mantile Giuditta Section on Developmental Genetics, Heritable Disorders Branch, Building 10, Room 9S241, NIH, 20892-1830, Bethesda, MD, U.S.A aut Bazzo Renzo IRBM P. Angeletti, Spa., Via Pontina km 30.600, I-00040, Pomezia, Rome, Italy aut Miele Lucio Section on Developmental Genetics, Heritable Disorders Branch, Building 10, Room 9S241, NIH, 20892-1830, Bethesda, MD, U.S.A aut Paolillo Livio Department of Chemistry, University of Naples 'Federico II‘, Via Mezzocannone 4, I-80100, Naples, Italy aut Mukherjee Anil Section on Developmental Genetics, Heritable Disorders Branch, Building 10, Room 9S241, NIH, 20892-1830, Bethesda, MD, U.S.A aut Barbato Gaetano IRBM P. Angeletti, Spa., Via Pontina km 30.600, I-00040, Pomezia, Rome, Italy aut Journal of Biomolecular NMR Kluwer Academic Publishers 9/1(1997-01-01), 35-46 0925-2738 9:1<35 1997 9 10858 https://doi.org/10.1023/A:1018619501038 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1018619501038 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Carlomagno Teresa Department of Chemistry, University of Naples 'Federico II‘, Via Mezzocannone 4, I-80100, Naples, Italy aut NATIONALLICENCE 700 1- Mantile Giuditta Section on Developmental Genetics, Heritable Disorders Branch, Building 10, Room 9S241, NIH, 20892-1830, Bethesda, MD, U.S.A aut NATIONALLICENCE 700 1- Bazzo Renzo IRBM P. Angeletti, Spa., Via Pontina km 30.600, I-00040, Pomezia, Rome, Italy aut NATIONALLICENCE 700 1- Miele Lucio Section on Developmental Genetics, Heritable Disorders Branch, Building 10, Room 9S241, NIH, 20892-1830, Bethesda, MD, U.S.A aut NATIONALLICENCE 700 1- Paolillo Livio Department of Chemistry, University of Naples 'Federico II‘, Via Mezzocannone 4, I-80100, Naples, Italy aut NATIONALLICENCE 700 1- Mukherjee Anil Section on Developmental Genetics, Heritable Disorders Branch, Building 10, Room 9S241, NIH, 20892-1830, Bethesda, MD, U.S.A aut NATIONALLICENCE 700 1- Barbato Gaetano IRBM P. Angeletti, Spa., Via Pontina km 30.600, I-00040, Pomezia, Rome, Italy aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Kluwer Academic Publishers 9/1(1997-01-01), 35-46 0925-2738 9:1<35 1997 9 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer