Macromolecular crystallography and structural biology databases at NIST - National Institute of Standards and Technology

Journal of Research of the National Institute of Standards and Technology, Nov, 2001 by Gary L. Gilliland

In the late 1970s, macromolecular crystallography at NIST began with collaboration between NIST and NIH to establish a single-crystal neutron diffractometer. This instrument was constructed and employed to solve a number of crystal structures: bovine ribonuclease A, bovine-ribonuclease-uridine vanadate complex, and porcine insulin. In the mid 1980s a Biomolecular Structure Group was created establishing NIST capabilities in biomolecular singe-crystal x-ray diffraction. The group worked on a variety of structural problems until joining the NIST/UMBI Center for Advanced Research in Biotechnology (CARB) in 1987. Crystallographic studies at CARB were then focused on protein engineering efforts that included among others chymosin, subtilisin BPN'. interleukin 1[beta], and glutathione Stransferase. Recently, the structural biology efforts have centered on enzymes in the chorismate metabolic pathways involved in amino acid biosynthesis and in structural genomics that involves determining the structures of "hypotheti cal" proteins to aid in assigning function. In addition to crystallographic studies, structural biology database activities began with the formal establishment of the Biological Macromolecule Crystallization Database in 1989. Later, in 1997, NIST in partnership with Rutgers and UCSD formed the Research Collaboratory for Structural Bioinformatics that successfully acquired the Protein Data Bank. The NIST efforts in these activities have focused on data uniformity, establishing and maintaining the physical archive, and working with the NMR community.

Key words: macromolecular crystallography; neutron crystallography; protein crystallography; proteins; structural biology databases; x-ray crystallography.

1. Introduction

structural biology studies began at NIST in the late 1970s when it was recognized that neutron diffraction methods could be used to obtain novel information about the atomic structure of macromolecules, especially in its ability to elucidate hydrogen atom positions. NIH and NBS established a collaborative arrangement to develop macromolecular neutron crystallographic capabilities. Early work by Dr. John Norvell and later by Dr. Alexander Wlodawer resulted in the development and implementation of a neutron diffractometer with a linear detector specifically designed for collecting diffraction data from crystals of biological macromolecules (1). The availability of the neutron diffractometer led to the determination of a number of protein structures. The requirements for these studies included protein crystals with relatively small unit cells, because of the diffraction data resolution requirements of the linear detector, and extremely large crystals (several millimeters in each dimension), because of the weak flux of the neutron beam (2).

In the mid 1980s the Biomolecular Structure Group was created in the Chemical Thermodynamics Division of the Center for Chemical Physics at NBS. Dr. Alexander Wlodawer who had been involved in establishing the NBS neutron diffractometer led this effort. This group established the first single-crystal macromolecular x-ray crystallographic laboratory at NBS. A number of important crystallographic studies were undertaken, many of which were completed prior to the incorporation of the group into the Center for Advanced Research in Biotechnology (CARB). This Center was established in the late 1980s when NIST began a long-term partnership with the University of Maryland. GARB was subsequently included as one of the Centers of the University of Maryland Biotechnology Institute (UMBI). The NIST crystallographic studies were focused on protein engineering. A number of productive structural investigations of proteins of industrial importance were undertaken, e.g., subtilisin (3), chymosin (4), and interleukin-1[beta] ( 5). As the CARB structural biology program matured, numerous other projects developed and were completed that have made significant contributions to understanding of how protein structure relates to function. Investigations of glutathione S-transferase (6), hemoglobin (7), uracil N-glycosylase (8), chorismate metabolism enzymes (9), and hypothetical protein targets associated with a structural genomics program (10) are representative of these efforts.

In addition to macromolecular crystallography, NIST staff members have been involved in the development and implementation of two important structural biology databases, the NIST/CARB Biological Macromolecule Crystallization Database (11) and the Protein Data Bank (12). These efforts have involved collaborations with other laboratories and have been and continue to be important resources for the structural biology and other research communities.

The NBS/NIST structural biology efforts have been extremely productive over the years and have involved many NBS/NIST and CARB scientists, their collaborators, and numerous guest workers. Below the NBS/NIST history and achievements in structural biology are highlighted.

2. Neutron Diffraction Studies