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Molecular Targets and Strategies for Inhibition of the Bacterial Type III Secretion System (T3SS); Inhibitors Directly Binding to T3SS Components

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Molecular Targets and Strategies for Inhibition of the Bacterial Type III Secretion System (T3SS); Inhibitors Directly Binding to T3SS Components biomolecules Review Molecular Targets and Strategies for Inhibition of the Bacterial Type III Secretion System (T3SS); Inhibitors Directly Binding to T3SS Components Julia A. Hotinger , Heather A. Pendergrass and Aaron E. May * Department of Medicinal Chemistry, School of Pharmacy, Virginia Commonwealth University, Richmond, VA 23219, USA; [email protected] (J.A.H.); [email protected] (H.A.P.) * Correspondence: [email protected] Abstract: The type III secretion system (T3SS) is a virulence apparatus used by many Gram-negative pathogenic bacteria to cause infections. Pathogens utilizing a T3SS are responsible for millions of infections yearly. Since many T3SS knockout strains are incapable of causing systemic infection, the T3SS has emerged as an attractive anti-virulence target for therapeutic design. The T3SS is a multiprotein molecular syringe that enables pathogens to inject effector proteins into host cells. These effectors modify host cell mechanisms in a variety of ways beneficial to the pathogen. Due to the T3SS’s complex nature, there are numerous ways in which it can be targeted. This review will be focused on the direct targeting of components of the T3SS, including the needle, translocon, basal body, sorting platform, and effector proteins. Inhibitors will be considered a direct inhibitor if they have a binding partner that is a T3SS component, regardless of the inhibitory effect being structural or functional. Citation: Hotinger, J.A.; Pendergrass, Keywords: H.A.; May, A.E. Molecular Targets type III secretion system; anti-virulence; inhibition; antibacterials and Strategies for Inhibition of the Bacterial Type III Secretion System (T3SS); Inhibitors Directly Binding to T3SS Components. Biomolecules 2021, 1. Introduction 11, 316. https://doi.org/10.3390/ The discovery and use of antibiotics have contributed to the overall increase in life biom11020316 expectancy throughout the world [1]. Notwithstanding the great success of antibiotic therapy, the over-prescription and misuse of broad-spectrum antibiotics have led to new Academic Editor: Wiesław Swi˛etnicki´ challenges. Resistance to antibiotics threatens to make our current bacterial infection treat- ments ineffective [2]. One solution to the resistance problem is the use of anti-virulence Received: 4 January 2021 therapeutics. This involves targeting the ability of pathogens to cause and sustain in- Accepted: 17 February 2021 fections. Anti-virulence drugs would stop or slow infection while exhibiting minimal Published: 19 February 2021 selective pressure [3]. One notable anti-virulence target is the type III secretion system (T3SS), a virulence factor used by pathogenic Gram-negative bacteria [4,5]. Some human Publisher’s Note: MDPI stays neutral pathogens that use T3SSs include, but are not limited to, Bordetella spp. [6], Burkholde- with regard to jurisdictional claims in ria spp. [7], Chlamydia spp. [8], enteropathogenic and enterohemorrhagic Escherichia coli published maps and institutional affil- (EPEC and EHEC, respectively) [9–11], Pseudomonas aeruginosa [4], Salmonella spp. [5], iations. Shigella spp. [12], Vibrio cholerae [13], and Yersinia spp. [14,15]. Pathogens targeting non- mammalian (Aeromonas spp. [16], Edwardsiella spp. [17], Vibrio spp. [18]) and plant hosts (Erwinia spp. [19], Pseudomonas syringae [20,21], Ralstonia spp. [20], Xanthomonas spp. [22]) are also found in nature. Given the conserved nature of the T3SS among pathogens, this Copyright: © 2021 by the authors. target lends itself to the possible development of therapeutics capable of treating multiple Licensee MDPI, Basel, Switzerland. infections, whether in human medical care, livestock upkeep, or agriculture. This article is an open access article The T3SS is a protein complex that secretes proteins directly from the cytosol of the distributed under the terms and bacterial pathogen into the host cell via a tube spanning the bacterial and host membranes. conditions of the Creative Commons Attribution (CC BY) license (https:// Any protein that has been secreted by the T3SS is considered an effector protein. Some creativecommons.org/licenses/by/ include the T3SS needle and translocon components in this distinction due to their sec- 4.0/). ondary effects as virulence factors or an outdated convention [23–26], but in this review Biomolecules 2021, 11, 316. https://doi.org/10.3390/biom11020316 https://www.mdpi.com/journal/biomolecules Biomolecules 2021, 11, 316 2 of 35 Biomolecules 2021, 11, 316 2 of 35 include the T3SS needle and translocon components in this distinction due to their sec- ondary effects as virulence factors or an outdated convention [23–26], but in this review effector proteinseffector will be proteins considered will beany considered protein secreted any protein by the secreted completed by the T3SS. completed These T3SS. These molecular syringe-likemolecular injections syringe-like have injectionsgiven the haveT3SS giventhe nickname the T3SS “the the type nickname III injecti- “the type III injec- some” [2,27–31].tisome” The structure [2,27–31 ].of Thethe injectis structureome of can the be injectisome broken down can into be broken five major down re- into five major gions: the translocon,regions: needle, the translocon, basal body, needle, export basal apparatus, body, export and apparatus, cytoplasmic and complex cytoplasmic complex (Figure 1). The(Figure translocon1). The (Figure translocon 1, dark (Figure green)1 ,is dark a protein green) hexamer is a protein pore hexamer that is formed pore that is formed in the host cell inmembrane the host cell that membrane binds to the that needle binds tip to and the needleallows tipsecreted and allows proteins secreted to pass proteins to pass into the host [32–35].into the The host needle [32–35 tip]. Theprotein needle (Figure tip protein 1, light (Figure green)1 is, lightalso green)considered is also the considered the translocon componenttranslocon by componentsome but will by be some considered but will bepart considered of the needle part in of this the review. needle in this review. The needle itselfThe is needlecomposed itself of is helical composed monomers of helical (Figure monomers 1, yellow) (Figure that1 form, yellow) a tube-like that form a tube-like structure after structurepolymerization after polymerization [31]. The needle [ 31di].ffers The in needlelength from differs species in length to strain from de- species to strain pending on thedepending pathogen’s on target the pathogen’shost [36]. These target variations host [36]. will These be discussed variations in will the be nee- discussed in the dle inhibition section.needle inhibition section. Figure 1. TheFigure general 1. The structure general of structure the T3SS. Leftof the: Cross-section T3SS. Left: Cross-section of a prototypical of a T3SSprototypical with color-coded T3SS with and color- labeled sections. Right: Tablecoded of unified and labeled names ofsections. T3SS structural Right: Table components of unified by names section of [ T3SS23]. structural components by sec- tion [23]. The basal body is made of a dual ring system with the inner ring (Figure1, light purple) The basalspanning body is made the inner of a bacterialdual ring membrane system with (IM) the and inner the ring outer (Figure ring (Figure 1, light1, dark purple) purple) spanningspanning the inner the bact peptidoglycanerial membrane layer (IM) (PG) and and the outer outer bacterial ring (Figure membrane 1, dark (OM), anchoring purple) spanningthe the needle peptidoglycan to the bacterial layer (PG) cell surfaceand outer [37 ].bacterial The export membrane apparatus (OM), (Figure an- 1, orange) choring the needleworks to with the bacterial the cytoplasmic cell surfac complexe [37]. (FigureThe export1, blue) apparatus to sort, guide,(Figure and 1, or- power secretion, ange) works withand the together cytoplasmic are called complex the sorting (Figure platform 1, blue) [to38 sort,]. The guide, cytoplasmic and power complex se- contains an cretion, and togetherATPase are (Figure called1 , the dark sorting blue) toplatform power secretion.[38]. The Thecytoplasmic sorting platform complex also con- functions as the tains an ATPaserecognition (Figure 1, domaindark blue) for to effectors. power secretion. It receives The them sorting from platform chaperone also proteins func- and unfolds tions as the recognitionthe effectors domain for secretion for effectors. as folded It receives proteins them are from too large chaperone to enter proteins the needle (~2.5 nm and unfolds theinner effectors diameter) for secretion[39–41] .as Oncefolded the proteins secreted are proteins too large pass to enter into the the needle host cell, they elicit (~2.5 nm innerspecific diameter) responses [39–41]. to Once reprogram the secr theeted host’s proteins machinery pass into to enablethe host colonization. cell, they In addition elicit specific responsesto reprogramming to reprogram (Salmonella the hostspp.,’s machineryShigella tospp.) enable [29, 30colonization.], some effectors In ad- kill the target dition to reprogrammingcells (Yersinia (Salmonellaspp., Pseudomonas spp., Shigellaspp.) spp.) [31 ,[29,30],39]. Individual some effectors pathogens kill the can tar- have upwards of get cells (Yersinia30 uniquespp., Pseudomonas genes for effectors spp.) [31,39]. [42–44 Individual]. pathogens can have upwards of 30 unique genes Manyfor effectors T3SS knockout[42–44]. strains have attenuated or eliminated virulence, making the Many T3SST3SS knockout an attractive strains anti-virulence have attenuat
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