Synchrotron-based macromolecular crystallography module for an undergraduate biochemistry laboratory course

Abstract

This paper describes the introduction of synchrotron-based macromolecular crystallography (MX) into an undergraduate laboratory class. An introductory 2 week experimental module on MX, consisting of four laboratory sessions and two classroom lectures, was incorporated into a senior-level biochemistry class focused on a survey of biochemical techniques, including the experimental characterization of proteins. Students purified recombinant protein samples, set up crystallization plates and flash-cooled crystals for shipping to a synchrotron. Students then collected X-ray diffraction data sets from their crystals the remote interface of the Molecular Biology Consortium beamline (4.2.2) at the Advanced Light Source in Berkeley, CA, USA. Processed diffraction data sets were transferred back to the laboratory and used in conjunction with partial protein models provided to the students for refinement and model building. The laboratory component was supplemented by up to 2 h of lectures by faculty with expertise in MX. This module can be easily adapted for implementation into other similar undergraduate classes, assuming the availability of local crystallographic expertise and access to remote data collection at a synchrotron source.

An All‐Inclusive and Straightway Laboratory Activity to Solve the Three‐Dimensional Crystal Structure of a Protein

“…Usually, these reports deal only with the individual steps, leaving incomplete the workflow performed by protein crystallographers. Only a few reports encompass all these activities in a single paper ; however, from our point of view, they require previous X‐ray crystallographic expertise.…”

X-ray Crystallography Analysis of Complexes Synthesized with Tris(2-pyridylmethyl)amine: A Laboratory Experiment for Undergraduate Students Integrating Interdisciplinary Concepts and Techniques

“…Nonetheless, X-ray crystallography is one of the most powerful analysis techniques currently available and is responsible for major advancements in several disciplines leading to Nobel Prizes awarded to 49 people for their work employing this technique . Despite its complicated underlying theory and instrument acquisition costs, institutions have successfully incorporated X-ray crystallography in their undergraduate curriculum using approaches such as Internet-facilitated remote diffractometer control, and utilizing case study education methods . Over 40 papers focused on X-ray crystallography have been published in this Journal with several emphasizing the importance of developing new experiment modules for undergraduate students to expose them to this technique. − …”

Connecting Key Concepts with Student Experience: Introducing Small-Molecule Crystallography to Chemistry Undergraduates Using a Flexible Laboratory Module

“…Atom connectivity, relative and absolute stereochemistry, and packing information can be determined in a single experiment more definitively than through other common techniques such as NMR, IR, or GC-MS. Thanks to the widespread availability of modern technology and software, currently even chemistry students and postdoctoral researchers who are not trained in the details of diffraction physics are able to routinely perform small-molecule single-crystal X-ray structure analysis. − Despite its complicated underlying theory and instrument acquisition costs, X-ray crystallography can be incorporated into an undergraduate curriculum via varied approaches, − including national laboratory visits and Internet-facilitated remote data collection. , As an alternative to dedicated X-ray crystallography courses designed for upper-level undergraduate and graduate students, ,, this paper describes a 3 h crystallography laboratory module for implementation in undergraduate experimental chemistry courses, including inorganic, organic, and physical chemistry, as well as for outreach programs such as interinstitutional visits . The module provides an opportunity for course instructors who are not familiar with crystallography, or for colleges that lack access to single-crystal X-ray diffractometers, to integrate X-ray crystallography into their undergraduate curriculum with a half-day trip.…”

“…2−4 Despite its complicated underlying theory and instrument acquisition costs, X-ray crystallography can be incorporated into an undergraduate curriculum via varied approaches, 5−16 including national laboratory visits 14 and Internet-facilitated remote data collection. 15,16 As an alternative to dedicated Xray crystallography courses designed for upper-level undergraduate and graduate students, 14,17,18 this paper describes a 3 h crystallography laboratory module for implementation in undergraduate experimental chemistry courses, including inorganic, organic, and physical chemistry, as well as for outreach programs such as interinstitutional visits. 19 The module provides an opportunity for course instructors who are not familiar with crystallography, or for colleges that lack access to single-crystal X-ray diffractometers, to integrate X-ray crystallography into their undergraduate curriculum with a half-day trip.…”