Abstract
Sec2p is an essential yeast gene and is part of the cell polarization process that leads to budding. The Nterminal domain of sec2p (Sec2pN) the guanine-nucleotide exchange factor for sec4p has been expressed in Escherichia coli, purified, and crystallized. Crystals belong to the space group P21 with unit cell dimensions 178.1 x 98.4 x 180.0 Å,β = 91.7°, and diffract synchrotron-generated X-rays to better than 3.6 Å resolution. Pseudo-precession plots reveal a Laue symmetry of 2/m, corresponding to the aforementioned space group, and unusual weak diffraction in the ∼5-7 Å resolution range. The Matthews number calculations for a typical crystal density suggest a range of 28 to 64 molecules per asymmetric unit. Self-rotation and native Patterson calculations demonstrate a pure helical array of protein subunits. Based on the X-ray diffraction data analysis and amino-acid sequence alignments, the paper presents a hypothetical model of the exchange domain of sec2p as a pair of coiled-coil helices that binds to sec4p and facilitates nucleotide disassociation.
Keywords: Pseudo-precession plot, Laue symmetry, self-rotation function, non-crystallographic symmetry, coiled-coil, nucleotide exchange factor
Protein & Peptide Letters
Title: Structural Insights into the Exchange Domain of Sec2p: Expression, Purification,Crystallization, and Preliminary X-Ray Diffraction Data Analysis
Volume: 14 Issue: 3
Author(s): Harindarpal S. Gill
Affiliation:
Keywords: Pseudo-precession plot, Laue symmetry, self-rotation function, non-crystallographic symmetry, coiled-coil, nucleotide exchange factor
Abstract: Sec2p is an essential yeast gene and is part of the cell polarization process that leads to budding. The Nterminal domain of sec2p (Sec2pN) the guanine-nucleotide exchange factor for sec4p has been expressed in Escherichia coli, purified, and crystallized. Crystals belong to the space group P21 with unit cell dimensions 178.1 x 98.4 x 180.0 Å,β = 91.7°, and diffract synchrotron-generated X-rays to better than 3.6 Å resolution. Pseudo-precession plots reveal a Laue symmetry of 2/m, corresponding to the aforementioned space group, and unusual weak diffraction in the ∼5-7 Å resolution range. The Matthews number calculations for a typical crystal density suggest a range of 28 to 64 molecules per asymmetric unit. Self-rotation and native Patterson calculations demonstrate a pure helical array of protein subunits. Based on the X-ray diffraction data analysis and amino-acid sequence alignments, the paper presents a hypothetical model of the exchange domain of sec2p as a pair of coiled-coil helices that binds to sec4p and facilitates nucleotide disassociation.
Export Options
About this article
Cite this article as:
Gill S. Harindarpal, Structural Insights into the Exchange Domain of Sec2p: Expression, Purification,Crystallization, and Preliminary X-Ray Diffraction Data Analysis, Protein & Peptide Letters 2007; 14 (3) . https://dx.doi.org/10.2174/092986607780090892
DOI https://dx.doi.org/10.2174/092986607780090892 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Quantitative Structure-Activity Relationship Studies: Understanding the Mechanism of Tyrosine Kinase Inhibition
Current Enzyme Inhibition Potential Role of <i>In Vitro-In Vivo</i> Correlations (IVIVC) for the Development of Plant-Derived Anticancer Drugs
Current Drug Targets STAT3: A Molecular Target for Cancer Whose Time Has Come
Current Signal Transduction Therapy Application of Bioinformatics for the Search of Novel Anti-Viral Therapies: Rational Design of Anti-Herpes Agents
Current Bioinformatics AIDS Related Viruses, their Association with Leukemia, and Raf Signaling
Current HIV Research Antitumor and Antiviral Activity of Pentacyclic Triterpenes
Mini-Reviews in Organic Chemistry Recent Advances in Targeted Anti-Vasculature Therapy: The Neuroblastoma Model
Current Drug Targets Exploring siRNA Umpired Nanogels: A Tale of Barrier Combating Carrier
Current Pharmaceutical Design Withdrawal Notice: Recent Developments in Anti-Cancer Activity of Compounds Containing the Thioether Group
Anti-Cancer Agents in Medicinal Chemistry From the Design to the Clinical Application of Thromboxane Modulators
Current Pharmaceutical Design Unraveling Potential Candidate Targets Associated with Expression of p16<sup>INK4a</sup> or p16 Truncated Fragment by Comparative Proteomics Analysis
Current Proteomics CycloSal-Pronucleotides of Brivudine Monophosphate - Highly Active Antiviral Agents
Current Medicinal Chemistry - Anti-Infective Agents Preventive and Therapeutic Effects of the Retinoid X Receptor Agonist Bexarotene on Tumors
Current Drug Metabolism Understanding Abnormal c-JNK/p38MAPK Signaling in Amyotrophic Lateral Sclerosis: Potential Drug Targets and Influences on Neurological Disorders
CNS & Neurological Disorders - Drug Targets How Do Microtubule-Targeted Drugs Work? An Overview
Current Cancer Drug Targets Melatonin Induces Apoptosis and Inhibits the Proliferation of Cancer Cells via Reactive Oxygen Species-mediated MAPK and mTOR Pathways
Clinical Cancer Drugs Docking Prediction for Luteolin Inhibiting TNF-α and NF-κB Pathway
Letters in Drug Design & Discovery Novel Drug Therapies for Fertility Preservation in Men Undergoing Chemotherapy: Clinical Relevance of Protector Agents
Current Medicinal Chemistry Need to Think Outside Organ-based Diagnosis to Molecular Diagnostics
Applied Clinical Research, Clinical Trials and Regulatory Affairs Wide-Ranging Genomic Effects of Plasticisers and Related Compounds
Current Drug Metabolism