Protein Targeting to Starch Synthase 1, a Functional Protein of Starch Biosynthesis in Wheat (Triticum Aestivum L.)

Abstract In cereal endosperm, native starch comprising amylose and amylopectin is synthesized by the coordinated activities of several pathway enzymes. Amylose in starch influences its physio-chemical properties resulting in several human health benefits. The Granule-Bound Starch Synthase I (GBSSI) is the most abundant starch-associated protein. GBSSI lacks dedicated Carbohydrate-binding module (CBM). Previously, Protein Targeting Starch Synthase 1 (PTST1) was identified as a crucial protein for the localization of GBSSI to the starch granules in Arabidopsis. The function of its homologous protein in the wheat endosperm is not known. In this study, TaPTST1, an AtPTST1 homolog, containing a CBM and a coiled-coil domain was identified in wheat. Protein-coding nucleotide sequence of TaPTST1 from Indian wheat variety ‘C 306’ was cloned and characterized. Homology modelling and molecular docking suggested the potential interaction of TaPTST1 with glucans and GBSSI. The TaPTST1 expression was higher in wheat grain than the other tissues, suggesting a grain-specific function. In vitro binding assays demonstrated different binding affinities of TaPTST1 for native starch, amylose, and amylopectin. Furthermore, the immunoaffinity pull-down assay revealed that TaPTST1 directly interacts with GBSSI, and the interaction is mediated by a coiled-coil domain. The direct protein-protein interaction was further confirmed by bimolecular fluorescence complementation assay (BiFC) in planta. Based on our findings we postulate a functional role for TaPTST1 in starch metabolism by targeting GBSSI to starch granules in wheat endosperm.

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PROTEIN TARGETING TO STARCH Is Required for Localising GRANULE-BOUND STARCH SYNTHASE to Starch Granules and for Normal Amylose Synthesis in Arabidopsis

PLoS Biology ◽

10.1371/journal.pbio.1002080 ◽

2015 ◽

Vol 13 (2) ◽

pp. e1002080 ◽

Cited By ~ 60

Author(s):

David Seung ◽

Sebastian Soyk ◽

Mario Coiro ◽

Benjamin A. Maier ◽

Simona Eicke ◽

...

Keyword(s):

Protein Targeting ◽

Starch Synthase ◽

Starch Granules ◽

Granule Bound Starch Synthase

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Survival Beyond the Perinatal Period Expands the Phenotypes Caused by Mutations in GLE1

10.1101/124172 ◽

2017 ◽

Author(s):

Edith Said ◽

Jessica X. Chong ◽

Maja Hempel ◽

Jonas Denecke ◽

Paul Soler ◽

...

Keyword(s):

Protein Targeting ◽

Coiled Coil ◽

Perinatal Period ◽

Severe Form ◽

Anterior Horn ◽

Anterior Horn Cell ◽

Cell Disease ◽

Feeding Difficulties ◽

Coiled Coil Domain ◽

Anterior Horn Cell Disease

AbstractMutations in GLE1 underlie Lethal Congenital Contracture syndrome (LCCS1) and Lethal Arthrogryposis with Anterior Horn Cell Disease (LAAHD). Both LCCS1 and LAAHD are characterized by reduced fetal movements, congenital contractures, and a severe form of motor neuron disease that results in fetal death or death in the perinatal period, respectively. Via trio-exome sequencing, we identified bi-allelic mutations in GLE1 in two unrelated individuals with motor delays, feeding difficulties and respiratory insufficiency who survived beyond the perinatal period. Each affected child had missense variants predicted to result in amino acid substitutions near the C-terminus of GLE1 that are predicted to disrupt protein-protein interaction or GLE1 protein targeting. We hypothesize that mutations that preserve function of the coiled-coil domain of GLE1 cause LAAHD whereas mutations that abolish the function of the coiled-coil domain cause LCCS1. The phenotype of LAAHD is now expanded to include multiple individuals surviving into childhood suggesting that LAAHD is a misnomer and should be re-named Arthrogryposis with Anterior Horn Cell Disease (AAHD). Too few cases have been reported to identify significant genotype-phenotype relationships, but given that perinatal lethality in AAHD typically resulted from respiratory failure, it is possible that early or aggressive airway management such as early tracheostomy and ventilation may enable survival beyond the perinatal period.

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Faculty Opinions recommendation of PROTEIN TARGETING TO STARCH is required for localising GRANULE-BOUND STARCH SYNTHASE to starch granules and for normal amylose synthesis in Arabidopsis.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.725366521.793504598 ◽

2015 ◽

Author(s):

Ulf-Ingo Flugge

Keyword(s):

Protein Targeting ◽

Starch Synthase ◽

Starch Granules ◽

Granule Bound Starch Synthase

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TMCC3 localizes at the three-way junctions for the proper tubular network of the endoplasmic reticulum

Biochemical Journal ◽

10.1042/bcj20190359 ◽

2019 ◽

Vol 476 (21) ◽

pp. 3241-3260

Author(s):

Sindhu Wisesa ◽

Yasunori Yamamoto ◽

Toshiaki Sakisaka

Keyword(s):

Endoplasmic Reticulum ◽

Membrane Proteins ◽

Membrane Protein ◽

Mammalian Cells ◽

Coiled Coil ◽

Transmembrane Domains ◽

Domain Family ◽

Coiled Coil Domain ◽

U2os Cells ◽

Er Membrane

The tubular network of the endoplasmic reticulum (ER) is formed by connecting ER tubules through three-way junctions. Two classes of the conserved ER membrane proteins, atlastins and lunapark, have been shown to reside at the three-way junctions so far and be involved in the generation and stabilization of the three-way junctions. In this study, we report TMCC3 (transmembrane and coiled-coil domain family 3), a member of the TEX28 family, as another ER membrane protein that resides at the three-way junctions in mammalian cells. When the TEX28 family members were transfected into U2OS cells, TMCC3 specifically localized at the three-way junctions in the peripheral ER. TMCC3 bound to atlastins through the C-terminal transmembrane domains. A TMCC3 mutant lacking the N-terminal coiled-coil domain abolished localization to the three-way junctions, suggesting that TMCC3 localized independently of binding to atlastins. TMCC3 knockdown caused a decrease in the number of three-way junctions and expansion of ER sheets, leading to a reduction of the tubular ER network in U2OS cells. The TMCC3 knockdown phenotype was partially rescued by the overexpression of atlastin-2, suggesting that TMCC3 knockdown would decrease the activity of atlastins. These results indicate that TMCC3 localizes at the three-way junctions for the proper tubular ER network.

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Faculty Opinions recommendation of The Rad50 coiled-coil domain is indispensable for Mre11 complex functions.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.717967809.793467391 ◽

2012 ◽

Author(s):

John Tainer

Keyword(s):

Coiled Coil ◽

Coiled Coil Domain ◽

Mre11 Complex ◽

Complex Functions

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The coiled-coil domain of EspA is essential for the assembly of the type III secretion translocon on the surface of enteropathogenic Escherichia coli. Vol. 274 (1999) 35969–35974

Journal of Biological Chemistry ◽

10.1016/s0021-9258(20)67595-0 ◽

2005 ◽

Vol 280 (19) ◽

pp. 19436

Author(s):

Robin M. Delahay ◽

Stuart Knutton ◽

Robert K. Shaw ◽

Elizabeth L. Hartland ◽

Mark J. Pallen ◽

...

Keyword(s):

Escherichia Coli ◽

Type Iii Secretion ◽

Coiled Coil ◽

Enteropathogenic Escherichia Coli ◽

Type Iii ◽

Coiled Coil Domain

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The git5 Gβ and git11 Gγ Form an Atypical Gβγ Dimer Acting in the Fission Yeast Glucose/cAMP Pathway

Genetics ◽

10.1093/genetics/157.3.1159 ◽

2001 ◽

Vol 157 (3) ◽

pp. 1159-1168 ◽

Cited By ~ 5

Author(s):

Sheila Landry ◽

Charles S Hoffman

Keyword(s):

Fission Yeast ◽

Mammalian Cells ◽

Glucose Repression ◽

Coiled Coil ◽

Carboxy Terminus ◽

Amino Terminal ◽

Camp Pathway ◽

Coiled Coil Domain ◽

Mutational Activation ◽

Two Hybrid

AbstractFission yeast adenylate cyclase, like mammalian adenylate cyclases, is regulated by a heterotrimeric G protein. The gpa2 Gα and git5 Gβ are both required for glucose-triggered cAMP signaling. The git5 Gβ is a unique member of the Gβ family in that it lacks an amino-terminal coiled-coil domain shown to be essential for mammalian Gβ folding and interaction with Gγ subunits. Using a git5 bait in a two-hybrid screen, we identified the git11 Gγ gene. Co-immunoprecipitation studies confirm the composition of this Gβγ dimer. Cells deleted for git11 are defective in glucose repression of both fbp1 transcription and sexual development, resembling cells lacking either the gpa2 Gα or the git5 Gβ. Overexpression of the gpa2 Gα partially suppresses loss of either the git5 Gβ or the git11 Gγ, while mutational activation of the Gα fully suppresses loss of either Gβ or Gγ. Deletion of gpa2 (Gα), git5 (Gβ), or git11 (Gγ) confer quantitatively distinct effects on fbp1 repression, indicating that the gpa2 Gα subunit remains partially active in the absence of the Gβγ dimer and that the git5 Gβ subunit remains partially active in the absence of the git11 Gγ subunit. The addition of the CAAX box from the git11 Gγ to the carboxy-terminus of the git5 Gβ partially suppresses the loss of the Gγ. Thus the Gγ in this system is presumably required for localization of the Gβγ dimer but not for folding of the Gβ subunit. In mammalian cells, the essential roles of the Gβ amino-terminal coiled-coil domains and Gγ partners in Gβ folding may therefore reflect a mechanism used by cells that express multiple forms of both Gβ and Gγ subunits to regulate the composition and activity of its G proteins.

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Comparison of Morphological and Physicochemical Properties of a Floury Rice Variety upon Pre-Harvest Sprouting

Foods ◽

10.3390/foods10040746 ◽

2021 ◽

Vol 10 (4) ◽

pp. 746

Author(s):

Chae-Min Han ◽

Jong-Hee Shin ◽

Jung-Bae Kwon ◽

Jong-Soo Kim ◽

Jong-Gun Won ◽

...

Keyword(s):

Physicochemical Properties ◽

Rice Variety ◽

Wheat Variety ◽

Economic Loss ◽

Rice Grain ◽

Starch Granules ◽

Rice Varieties ◽

Starch Structure ◽

Gelatinization Properties

Pre-harvest sprouting (PHS) severely reduces rice grain yield, significantly affects grain quality, and leads to substantial economic loss. In this study, we aimed to characterize the physicochemical properties and processing quality of the Garumi 2 flour rice variety under PHS conditions and compare them with those of the Seolgaeng, Hangaru, Shingil, and Ilpum rice varieties and the Keumkang wheat variety. Analysis of the molecular structure of starch revealed uniform starch granules, increased proportions of short-chain amylopectin in DP 6–12 (51.0–55.3%), and enhanced crystallinity (30.7–35.7%) in rice varieties for flour compared with the Ilpum cooking rice variety. PHS significantly altered the starch structure and gelatinization properties of Garumi 2. It also caused surface pitting and roughness in Garumi 2 starch granules and decreased their crystallinity. Collectively, the findings of this study provide important novel insights into the effects of PHS on the physicochemical properties of Garumi 2 floury rice for flour.

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