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Title: Serial Femtosecond X-Ray Diffraction of HIV-1 Gag MA-IP6 Microcrystals at Ambient Temperature

Abstract

The Human immunodeficiency virus-1 (HIV-1) matrix (MA) domain is involved in the highly regulated assembly process of the virus particles that occur at the host cell’s plasma membrane. High-resolution structures of the MA domain determined using cryo X-ray crystallography have provided initial insights into the possible steps in the viral assembly process. However, these structural studies have relied on large and frozen crystals in order to reduce radiation damage caused by the intense X-rays. Here, we report the first X-ray free-electron laser (XFEL) study of the HIV-1 MA domain’s interaction with inositol hexaphosphate (IP6), a phospholipid headgroup mimic. We also describe the purification, characterization and microcrystallization of two MA crystal forms obtained in the presence of IP6. In addition, we describe the capabilities of serial femtosecond X-ray crystallography (SFX) using an XFEL to elucidate the diffraction data of MA-IP6 complex microcrystals in liquid suspension at ambient temperature. Two different microcrystal forms of the MA-IP6 complex both diffracted to beyond 3.5 Å resolution, demonstrating the feasibility of using SFX to study the complexes of MA domain of HIV-1 Gag polyprotein with IP6 at near-physiological temperatures. Further optimization of the experimental and data analysis procedures will lead to better understanding of themore » MA domain of HIV-1 Gag and IP6 interaction at high resolution and will provide basis for optimization of the lead compounds for efficient inhibition of the Gag protein recruitment to the plasma membrane prior to virion formation.« less

Authors:
 [1];  [2];  [2];  [2];  [3];  [3];  [4];  [4];  [4];  [2];  [2];  [3];  [3]; ORCiD logo [2]
  1. Science Farm Ltd., Kumamoto (Japan); Kumamoto Univ., Kumamoto (Japan); SLAC National Accelerator Lab., Menlo Park, CA (United States)
  2. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  3. Kumamoto Univ., Kumamoto (Japan)
  4. KEK/High Energy Accelerator Research Organization, Ibaraki (Japan)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1528887
Grant/Contract Number:  
AC02-76SF00515; 1231306
Resource Type:
Accepted Manuscript
Journal Name:
International Journal of Molecular Sciences (Online)
Additional Journal Information:
Journal Name: International Journal of Molecular Sciences (Online); Journal Volume: 20; Journal Issue: 7; Journal ID: ISSN 1422-0067
Publisher:
MDPI
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; 59 BASIC BIOLOGICAL SCIENCES; Serial Femtosecond X-ray crystallography; human immunodeficiency virus; matrix protein; inositol hexaphosphate; ambient temperature

Citation Formats

Ciftci, Halil I., Sierra, Raymond G., Yoon, Chun Hong, Su, Zhen, Tateishi, Hiroshi, Koga, Ryoko, Kotaro, Koiwai, Yumoto, Fumiaki, Senda, Toshiya, Liang, Mengling, Wakatsuki, Soichi, Otsuka, Masami, Fujita, Mikako, and DeMirci, Hasan. Serial Femtosecond X-Ray Diffraction of HIV-1 Gag MA-IP6 Microcrystals at Ambient Temperature. United States: N. p., 2019. Web. doi:10.3390/ijms20071675.
Ciftci, Halil I., Sierra, Raymond G., Yoon, Chun Hong, Su, Zhen, Tateishi, Hiroshi, Koga, Ryoko, Kotaro, Koiwai, Yumoto, Fumiaki, Senda, Toshiya, Liang, Mengling, Wakatsuki, Soichi, Otsuka, Masami, Fujita, Mikako, & DeMirci, Hasan. Serial Femtosecond X-Ray Diffraction of HIV-1 Gag MA-IP6 Microcrystals at Ambient Temperature. United States. doi:10.3390/ijms20071675.
Ciftci, Halil I., Sierra, Raymond G., Yoon, Chun Hong, Su, Zhen, Tateishi, Hiroshi, Koga, Ryoko, Kotaro, Koiwai, Yumoto, Fumiaki, Senda, Toshiya, Liang, Mengling, Wakatsuki, Soichi, Otsuka, Masami, Fujita, Mikako, and DeMirci, Hasan. Wed . "Serial Femtosecond X-Ray Diffraction of HIV-1 Gag MA-IP6 Microcrystals at Ambient Temperature". United States. doi:10.3390/ijms20071675. https://www.osti.gov/servlets/purl/1528887.
@article{osti_1528887,
title = {Serial Femtosecond X-Ray Diffraction of HIV-1 Gag MA-IP6 Microcrystals at Ambient Temperature},
author = {Ciftci, Halil I. and Sierra, Raymond G. and Yoon, Chun Hong and Su, Zhen and Tateishi, Hiroshi and Koga, Ryoko and Kotaro, Koiwai and Yumoto, Fumiaki and Senda, Toshiya and Liang, Mengling and Wakatsuki, Soichi and Otsuka, Masami and Fujita, Mikako and DeMirci, Hasan},
abstractNote = {The Human immunodeficiency virus-1 (HIV-1) matrix (MA) domain is involved in the highly regulated assembly process of the virus particles that occur at the host cell’s plasma membrane. High-resolution structures of the MA domain determined using cryo X-ray crystallography have provided initial insights into the possible steps in the viral assembly process. However, these structural studies have relied on large and frozen crystals in order to reduce radiation damage caused by the intense X-rays. Here, we report the first X-ray free-electron laser (XFEL) study of the HIV-1 MA domain’s interaction with inositol hexaphosphate (IP6), a phospholipid headgroup mimic. We also describe the purification, characterization and microcrystallization of two MA crystal forms obtained in the presence of IP6. In addition, we describe the capabilities of serial femtosecond X-ray crystallography (SFX) using an XFEL to elucidate the diffraction data of MA-IP6 complex microcrystals in liquid suspension at ambient temperature. Two different microcrystal forms of the MA-IP6 complex both diffracted to beyond 3.5 Å resolution, demonstrating the feasibility of using SFX to study the complexes of MA domain of HIV-1 Gag polyprotein with IP6 at near-physiological temperatures. Further optimization of the experimental and data analysis procedures will lead to better understanding of the MA domain of HIV-1 Gag and IP6 interaction at high resolution and will provide basis for optimization of the lead compounds for efficient inhibition of the Gag protein recruitment to the plasma membrane prior to virion formation.},
doi = {10.3390/ijms20071675},
journal = {International Journal of Molecular Sciences (Online)},
number = 7,
volume = 20,
place = {United States},
year = {2019},
month = {4}
}

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