[Isolation, analysis of amino acid sequence and crystallization of the extracellular ribonuclease Th1 from Trichoderma harzianum-01].
A procedure of large-scale isolation of homogeneous ribonuclease Th1 from cultural filtrates of Trichoderma harzianum with a yield over 50% has been developed. Three ion-exchange chromatographies on CM- and DEAE-cellulose gave 7500 fold purification of the protein with a specific activity of ca. 4500 U/mg. The RNase Th1 is shown to be a basic protein (pI 9.5) with Mr 10,747; it contains 106 amino acid residues (2 Asp, 6 Asn, 9 Thr, 12 Ser, 2 Glu, 1 Gln, 4 Pro, 16 Gly, 14 Ala, 4 Cys, 7 Val, 5 Ile, 2 Leu, 7 Tyr, 6 Phe, 2 His, 4 Lys, 3 Arg). The total amino acid sequence of RNase Th1 was determined and, on comparison with other guanyl-specific fungal RNases, showed a significant degree of homology, thus indicating probability of a common origin. By means of the equilibrium dialysis, crystals of RNase Th1 were obtained with the space group P3(2)21, a = b = 55.7, c = 80.1 A. A preliminary X-ray study of RNase Th1 was undertaken.
D000595:Amino Acid Sequence; D002848:Chromatography, DEAE-Cellulose; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D004722:Endoribonucleases / Q000302:isolation & purification*; D003904:Mitosporic Fungi / Q000201:enzymology*; D008969:Molecular Sequence Data; D006163:Ribonuclease T1 / Q000032:analysis / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet; D014242:Trichoderma / Q000201:enzymology*
D004722:Endoribonucleases; D006163:Ribonuclease T1
Vydelenie, analiz aminokislotnoĭ posledovatel'nosti i kristallizatsiia vnekletochnoĭ ribonukleazy Th1 griba Trichoderma harzianum-01.