scholarly journals The encapsulin from Thermatoga maritima is a flavoprotein with a symmetry matched ferritin-like cargo protein

2021 ◽  
Author(s):  
Benjamin J LaFrance ◽  
Caleb Cassidy-Amstutz ◽  
Robert J Nichols ◽  
Luke M Oltrogge ◽  
Eva Nogales ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Biochemical Analysis ◽  
Active Role ◽  
Structure And Function ◽  
Redox Active ◽  
Cargo Protein ◽  
Cargo Proteins ◽  
And Function ◽  
Flavin Binding ◽  
Unique Chemical

Bacterial nanocompartments, also known as encapsulins, are an emerging class of protein-based "organelles" found in bacteria and archaea. Encapsulins are virus-like icosahedral particles comprising a ~25-50 nm shell surrounding a specific cargo enzyme. Compartmentalization is thought to create a unique chemical environment to facilitate catalysis and isolate toxic intermediates. Many questions regarding nanocompartment structure-function remain unanswered, including how shell symmetry dictates cargo loading and to what extent the shell facilitates enzymatic activity. Here, we explore these questions using the model T. maritima nanocompartment known to encapsulate a redox-active ferritin-like protein. Biochemical analysis revealed the encapsulin shell to possess a flavin binding site located at the interface between capsomere subunits, suggesting the shell may play a direct and active role in the function of the encapsulated cargo. Furthermore, we used cryoEM to show that cargo proteins use a form of symmetry-matching to facilitate encapsulation and define stoichiometry. In the case of the T. maritima encapsulin, the decameric cargo protein with 5-fold symmetry preferentially binds to the pentameric-axis of the icosahedral shell. Taken together, these observations suggest the shell is not simply a passive barrier-it also plays a significant role in the structure and function of the cargo enzyme.

scholarly journals The encapsulin from Thermotoga maritima is a flavoprotein with a symmetry matched ferritin-like cargo protein

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Benjamin J. LaFrance ◽  
Caleb Cassidy-Amstutz ◽  
Robert J. Nichols ◽  
Luke M. Oltrogge ◽  
Eva Nogales ◽  
...  
Keyword(s):  
Biochemical Analysis ◽  
Thermotoga Maritima ◽  
Active Role ◽  
Structure And Function ◽  
Redox Active ◽  
Cargo Protein ◽  
Cargo Proteins ◽  
And Function ◽  
Flavin Binding ◽  
Unique Chemical

AbstractBacterial nanocompartments, also known as encapsulins, are an emerging class of protein-based ‘organelles’ found in bacteria and archaea. Encapsulins are virus-like icosahedral particles comprising a ~ 25–50 nm shell surrounding a specific cargo enzyme. Compartmentalization is thought to create a unique chemical environment to facilitate catalysis and isolate toxic intermediates. Many questions regarding nanocompartment structure–function remain unanswered, including how shell symmetry dictates cargo loading and to what extent the shell facilitates enzymatic activity. Here, we explore these questions using the model Thermotoga maritima nanocompartment known to encapsulate a redox-active ferritin-like protein. Biochemical analysis revealed the encapsulin shell to possess a flavin binding site located at the interface between capsomere subunits, suggesting the shell may play a direct and active role in the function of the encapsulated cargo. Furthermore, we used cryo-EM to show that cargo proteins use a form of symmetry-matching to facilitate encapsulation and define stoichiometry. In the case of the Thermotoga maritima encapsulin, the decameric cargo protein with fivefold symmetry preferentially binds to the pentameric-axis of the icosahedral shell. Taken together, these observations suggest the shell is not simply a passive barrier—it also plays a significant role in the structure and function of the cargo enzyme.

Photocatalytic redox reactions with metalloporphyrins

2020 ◽  
Vol 24 (01n03) ◽  
pp. 21-32 ◽  
Author(s):  
Shunichi Fukuzumi ◽  
Yong-Min Lee ◽  
Wonwoo Nam
Keyword(s):  
Hydrogen Evolution ◽  
Redox Reactions ◽  
Bond Formation ◽  
Visible Region ◽  
Photosynthetic Reaction Center ◽  
Structure And Function ◽  
Free Base ◽  
Redox Catalyst ◽  
Redox Active ◽  
And Function

Metalloporphyrinoids are utilized as efficient sensitizers and catalysts in photosynthesis and the reverse reaction that is respiration. Because metalloporphyrinoids show strong absorption in the visible region and redox active, metalloporphyrinoids are also suited as photoredox catalysts for photo-driven redox reactions using solar energy. In particular, metalloporphyrins are utilized as pivotal components to mimic the structure and function of the photosynthetic reaction center. Metalloporphyrins are used as photoredox catalysts for hydrogen evolution from electron and proton sources combining hydrogen evolution catalysts. Metalloporphyrins also act as thermal redox catalysts for photocatalytic reduction of CO2 with photoredox catalysts. Metalloporphyrins are also used as dual catalysts for a photoredox catalyst for oxygenation of substrates with H2O and a redox catalyst for O2 reduction when dioxygen is used as a two-electron oxidant and H2O as an oxygen source, both of which are the greenest reactants. Free base porphyrins can also be employed as promising photoredox catalysts for C–C bond formation reactions.

eIF2B, a mediator of general and gene-specific translational control

2005 ◽  
Vol 33 (6) ◽  
pp. 1487-1492 ◽  
Author(s):  
G.D. Pavitt
Keyword(s):  
Translational Control ◽  
Biochemical Analysis ◽  
Molecular Genetic ◽  
Structure And Function ◽  
Initiation Factor ◽  
Protein Synthesis Initiation ◽  
Vanishing White Matter Disease ◽  
And Function ◽  
Vanishing White Matter ◽  
Childhood Ataxia

eIF2B (eukaryotic initiation factor 2B) is a multisubunit protein that is required for protein synthesis initiation and its regulation in all eukaryotic cells. Mutations in eIF2B have also recently been found to cause a fatal human disease called CACH (childhood ataxia with central nervous system hypomyelination) or VWM (vanishing white matter disease). This review provides a general background to translation initiation and mechanisms known to control eIF2B function, before describing molecular genetic and biochemical analysis of eIF2B structure and function, integrating work from studies of the yeast and mammalian eIF2B proteins.

The haloarchaeal chromosome replication machinery

2009 ◽  
Vol 37 (1) ◽  
pp. 108-113 ◽  
Author(s):  
Stuart A. MacNeill
Keyword(s):  
Dna Replication ◽  
Replication Fork ◽  
Biochemical Analysis ◽  
Structure And Function ◽  
Haloferax Volcanii ◽  
Eukaryotic Cells ◽  
Replication Machinery ◽  
Chromosomal Dna ◽  
Biochemical Studies ◽  
And Function

The powerful combination of genetic and biochemical analysis has provided many key insights into the structure and function of the chromosomal DNA replication machineries of bacterial and eukaryotic cells. In contrast, in the archaea, biochemical studies have dominated, mainly due to the absence of efficient genetic systems for these organisms. This situation is changing, however, and, in this regard, the genetically tractable haloarchaea Haloferax volcanii and Halobacterium sp. NRC-1 are emerging as key models. In the present review, I give an overview of the components of the replication machinery in the haloarchaea, with particular emphasis on the protein factors presumed to travel with the replication fork.

scholarly journals N-linked Oligosaccharides Affect the Enzymatic Activity of CD39: Diverse Interactions between Seven N-linked Glycosylation Sites

2005 ◽  
Vol 16 (4) ◽  
pp. 1661-1672 ◽  
Author(s):  
James J. Wu ◽  
Lisa E. Choi ◽  
Guido Guidotti
Keyword(s):  
Enzymatic Activity ◽  
Surface Expression ◽  
Structure And Function ◽  
Surface Appearance ◽  
C Terminus ◽  
Glycosylation Sites ◽  
N Terminus ◽  
Membrane Bound ◽  
And Function ◽  
Diverse Interactions

Rat CD39, a membrane-bound ectonucleoside triphosphate diphosphohydrolase that hydrolyzes extracellular nucleoside tri- and diphosphates, has seven potential N-glycosylation sites at asparagine residues 73, 226, 291, 333, 375, 429, and 458. To determine their roles in the structure and function of CD39, we mutated these sites individually or in combination by replacing asparagine with serine or glutamine and analyzed the surface expression and the enzymatic activity of the mutants. The results indicate that rat CD39 can be glycosylated at all seven sites when expressed in COS7 cells. Glycosylation sites 73 at the N terminus, 333 in the middle, and 429 and 458 at the C terminus were principally required for cell surface appearance of enzymatically active CD39. Whereas deletion of these sites individually had modest effects on surface ATPase activity, some double deletions of these sites had major effects on both surface activity and expression. The importance of these N-glycosylation sites is recognizable in other members of the ectonucleoside triphosphate diphosphohydrolase family.

Biochemical Analysis of Chromatin Structure and Function UsingDrosophilaEmbryo Extracts

Methods ◽  
1997 ◽  
Vol 12 (1) ◽  
pp. 28-35 ◽  
Author(s):  
Thiemo A. Blank ◽  
Raphael Sandaltzopoulos ◽  
Peter B. Becker
Keyword(s):  
Chromatin Structure ◽  
Biochemical Analysis ◽  
Structure And Function ◽  
And Function

scholarly journals Exploring the influence of silver and lead on structure and function of xylanase: spectroscopic and calorimetric methods

2020 ◽  
Vol 9 (3) ◽  
pp. 182-190
Author(s):  
Mingyang Jing ◽  
Rui Tang ◽  
Guangye Han ◽  
Shansheng Zhang ◽  
Rutao Liu
Keyword(s):  
Enzymatic Activity ◽  
Molecular Level ◽  
Structure And Function ◽  
Titration Calorimetry ◽  
Soil Enzyme Activities ◽  
Hydrophobic Force ◽  
Negative Effect ◽  
Endogenous Fluorescence ◽  
Underlying Mechanisms ◽  
And Function

Abstract Soil contamination with heavy metal could induce the alteration of soil ecological environments, and soil enzyme activities are sensitive indicators for the soil toxicology. Xylanase is one of predominant soil enzymes related to carbon nitrogen cycle. In this work, we explored the underlying mechanisms for conformational and enzymatic activity alterations of xylanase after silver and lead exposure at molecular level with systematical measurements including multiple spectroscopic methods, isothermal titration calorimetry, and enzymatic activity. Both silver and lead could loosen and unfold the skeleton of xylanase with the quenching of endogenous fluorescence. Silver interacted with xylanase forming larger-size aggregations through Van der Waals forces and hydrogen bonding, while lead interacted with xylanase forming larger-size aggregations through hydrophobic force. Silver and lead induced an obvious loss (67.1 and 56.31%) of the xylanase enzymatic activity, but silver has a greater impact on xylanase than that of lead. The xylanase enzymatic activity significantly decreased due to the conformational alterations. The negative effect of silver exposure on xylanase structure and function was more prominent than that of lead.

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