Macromolecular crystallization: basics and advanced methodologies
JOURNAL OF THE IRANIAN CHEMICAL SOCIETY
Authors: Cheraghian Radi, Hamid; Hajipour-Verdom, Behnam; Molaabasi, Fatemeh
Abstract
For the first time, about 155 years ago and as a laboratory curiosity, German biologists observed the crystals of hemoglobin from worms and fishes. Since then, the crystallization of proteins, nucleic acids and big biological structures, like viruses, has been developed into a broad research field including several applications, for example in the drug discovery. This review is divided into four major sections. The first section addresses the specific physicochemical properties of biomolecular crystals accompanied by kinetics of supersaturation, nucleation and growth which are the three main steps required to achieve macromolecular crystals. Besides, various physical, chemical and biochemical parameters influencing the process of macromolecular crystallization are reviewed. The second part deals with classical approaches, such as vapor and batch diffusion methods, available to create macromolecular crystals. The third part overviews novel approaches including microgravity, cocrystallization, membrane-assisted crystallization and microfluidic array chips involved in more complicated techniques for growing macromolecular crystals and controlling their size and orientation. In the end, considering the very significant role of automation in providing biomolecules' crystals in recent years, we provided a brief explanation about robotics and their importance in developing high-throughput crystallization. RDC-NMR and SAXS/WAXS hybrid methods with the aim of obtaining structural information of complex macromolecular assemblies are also discussed.
Hantavirus Infection Is Inhibited by Griffithsin in Cell Culture
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
Authors: Shrivastava-Ranjan, Punya; Lo, Michael K.; Chatterjee, Payel; Flint, Mike; Nichol, Stuart T.; Montgomery, Joel M.; O'Keefe, Barry R.; Spiropoulou, Christina F.
Abstract
Andes virus (ANDV) and Sin Nombre virus (SNV), highly pathogenic hantaviruses, cause hantavirus pulmonary syndrome in the Americas. Currently no therapeutics are approved for use against these infections. Griffithsin (GRFT) is a high-mannose oligosaccharide-binding lectin currently being evaluated in phase I clinical trials as a topical microbicide for the prevention of human immunodeficiency virus (HIV-1) infection (ClinicalTrials.gov Identifiers: NCT04032717, NCT02875119) and has shown broad-spectrum in vivo activity against other viruses, including severe acute respiratory syndrome coronavirus, hepatitis C virus, Japanese encephalitis virus, and Nipah virus. In this study, we evaluated the in vitro antiviral activity of GRFT and its synthetic trimeric tandemer 3mGRFT against ANDV and SNV. Our results demonstrate that GRFT is a potent inhibitor of ANDV infection. GRFT inhibited entry of pseudo-particles typed with ANDV envelope glycoprotein into host cells, suggesting that it inhibits viral envelope protein function during entry. 3mGRFT is more potent than GRFT against ANDV and SNV infection. Our results warrant the testing of GRFT and 3mGRFT against ANDV infection in animal models.