Crystallization and preliminary x-ray data for a glycosylated form of bovine pancreatic ribonuclease.

B Rubin,V Carperos,E Kezar

doi:10.1016/s0021-9258(18)34215-7

Abstract

A glycosylated form of bovine pancreatic ribonuclease, ribonuclease B, has been crystallized in a form suitable for three-dimensional x-ray structure determination. Crystals grown from polyethylene glycol solutions display high resolution diffraction patterns indicating the orthorhombic space group P2(1)2(1)2(1) with a = 59.2 (+/- 0.1), b = 56.0 (+/- 0.1), and c = 81.0 (+/- 0.2). A, V = 2.7 x 10(5) A3. With 8 molecules/unit cell Vm = 2.28 A3/dalton. The systemically weak intensity of all reflections with their l indices odd suggests the likely alignment of a noncrystallographic 2-fold axis near the crystal c axis.

Highlights

Protein Purification-Purified ribonuclease B was obtained comsuitable for three-dimensional x-ray structuredeter- mercially fromSigma (TypeXII-B)and was purified further by mination
Crystalsgrown from polyethylene glycol so- affinity chromatography on concanavalin A-sepharose 4B (Pharmalutions display high resolution diffraction patterns in- cia) at 4 "C following the procedure of Baynes and Wold [13]
The combined ribonuclease fractions were dialyzed against 0.1 M Tris-HC1 at pH 7.3, and ment of a noncrystallographic 2-foaldxis near thecrys- concentrated by ultrafiltration on an Amicon YMlO membrane to a tal c axis

Summary

Introduction

Protein Purification-Purified ribonuclease B was obtained comsuitable for three-dimensional x-ray structuredeter- mercially fromSigma (TypeXII-B)and was purified further by mination. A glycosylated form of bovine pancreatic ribonuclease, ribonuclease B, has beencrystallizedinaform Crystalsgrown from polyethylene glycol so- affinity chromatography on concanavalin A-sepharose 4B (Pharmalutions display high resolution diffraction patterns in- cia) at 4 "C following the procedure of Baynes and Wold [13]. The systematically weak intensity of all reflections with their1 indices odd suggests the likely aligna - g~alactopyranoside was begun after washing the bound RNase B with only 2-4 column volumes of buffer.

Results

Conclusion