double domain swapping in bovine seminal rnase formation of distinct n- and c-swapped tetramers and multimers with increasing biological activities双域交换在牛的核糖核酸酶的形成不同的n - c-swapped四聚体和多聚体增加生物活性.pdfVIP

Double Domain Swapping in Bovine Seminal RNase: Formation of Distinct N- and C-swapped Tetramers and Multimers with Increasing Biological Activities 1 1 2 3 Giovanni Gotte *, Alexander Mahmoud Helmy , Carmine Ercole , Roberta Spadaccini , 4 1 2 Douglas V. Laurents , Massimo Donadelli , Delia Picone ` 1 Dipartimento di Scienze della Vita e della Riproduzione, Sezione di Chimica Biologica, Universita degli Studi di Verona, Verona, Italy, 2 Dipartimento di Scienze Chimiche, ` ` Universita degli Studi di Napoli ‘‘Federico II’’, Naples, Italy, 3 Dipartimento di Scienze Biologiche e Ambientali, Universita del Sannio, Benevento, Italy, 4 Instituto de ´ ´ Quımica Fısica ‘‘Rocasolano’’ (C.S.I.C.), Madrid, Spain Abstract Bovine seminal (BS) RNase, the unique natively dimeric member of the RNase super-family, represents a special case not only for its additional biological actions but also for the singular features of 3D domain swapping. The native enzyme is indeed a mixture of two isoforms: M = M, a dimer held together by two inter-subunit disulfide bonds, and MxM, 70% of the total, which, besides the two mentioned disulfides, is additionally stabilized by the swapping of its N-termini. When lyophilized from 40% acetic acid, BS-RNase oligomerizes as the super-family proto-type RNase A does. In this paper, we induced