Localization of Rac2 via the C terminus and aspartic acid 150 specifies superoxide generation, actin polarity and chemotaxis in neutrophils

2004 ◽  
Vol 5 (7) ◽  
pp. 744-751 ◽  
Author(s):  
Marie-Dominique Filippi ◽  
Chad E Harris ◽  
Jarek Meller ◽  
Yi Gu ◽  
Yi Zheng ◽  
...  
Keyword(s):  
Aspartic Acid ◽  
Superoxide Generation ◽  
C Terminus

scholarly journals Thermal Synthesis of Polypeptides from N-Butyloxycarbonyl Oligopeptides Containing Aspartyl Residue at C-Terminus

2017 ◽  
Vol 2017 ◽  
pp. 1-16
Author(s):  
Toratane Munegumi ◽  
Takafumi Yamada
Keyword(s):  
Aspartic Acid ◽  
Organic Synthesis ◽  
Prebiotic Chemistry ◽  
Average Molecular Weight ◽  
Carboxyl Terminus ◽  
Peptide Bonds ◽  
Thermal Synthesis ◽  
Thermal Reactions ◽  
C Terminus ◽  
Aspartyl Residue

The thermal reactions of amino acids have been investigated for pure organic synthesis, materials preparation in industry, and prebiotic chemistry. N-t-Butyloxycarbonyl aspartic acid (Boc-Asp) releases 2-butene and carbon dioxide upon heating without solvents. The resulting mixture of the free molten aspartic acid was dehydrated to give peptide bonds. This study describes the thermal reactions of N-t-butyloxycarbonyl peptides (Boc-Gly-L-Asp, Boc-L-Ala-L-Asp, Boc-L-Val-L-Asp, and Boc-Gly-Gly-L-Asp) having an aspartic residue at the carboxyl terminus. The peptides were deprotected upon heating at a constant temperature between 110 and 170°C for 1 to 24 h to afford polypeptides in which the average molecular weight reached 7800.

scholarly journals Export from the Endoplasmic Reticulum of AssembledN-Methyl-D-aspartic Acid Receptors Is Controlled by a Motif in the C Terminus of the NR2 Subunit

2004 ◽  
Vol 279 (28) ◽  
pp. 28903-28910 ◽  
Author(s):  
Lynda M. Hawkins ◽  
Kate Prybylowski ◽  
Kai Chang ◽  
Caroline Moussan ◽  
F. Anne Stephenson ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Aspartic Acid ◽  
C Terminus

scholarly journals Light-dependent THRUMIN1 phosphorylation regulates its association with actin filaments and 14-3-3 proteins

2021 ◽  
Author(s):  
Matthew E Dwyer ◽  
Roger P. Hangarter
Keyword(s):  
Aspartic Acid ◽  
Actin Filaments ◽  
Chloroplast Movement ◽  
Cell Surfaces ◽  
Intrinsically Disordered ◽  
C Terminus ◽  
Intrinsically Disordered Region ◽  
Chloroplast Movements ◽  
Cell Layers

Light-dependent chloroplast movements in leaf cells contribute to the optimization of photosynthesis. Low light conditions induce chloroplast accumulation along periclinal cell surfaces, providing greater access to the available light, whereas high light induces movement of chloroplasts to anticlinal cell surfaces providing photodamage protection and allowing more light to reach underlying cell layers. The THRUMIN1 protein is required for normal chloroplast movements in Arabidopsis thaliana and has been shown to localize at the plasma membrane and to undergo rapid light-dependent interactions with actin filaments through the N-terminal intrinsically disordered region. A predicted WASP-Homology 2 (WH2) domain was found in the intrinsically disordered region but mutations in this domain did not disrupt localization of THRUMIN1:YFP to actin filaments. A series of other protein truncations and site-directed mutations of known and putative phosphorylation sites indicated that a phosphomimetic mutation (serine to aspartic acid) at position 170 disrupted localization of THRUMIN1 with actin filaments. However, the phosphomimetic mutant rescued the thrumin1-2 mutant phenotype for chloroplast movement and raises questions about the role of THRUMIN1's interaction with actin. Mutation of serine 146 to aspartic acid also resulted in cytoplasmic localization of THRUMIN1:YFP in Nicotiana benthamiana. Mutations to a group of putative zinc-binding cysteine clusters implicates the C-terminus of THRUMIN1 in chloroplast movement. Phosphorylation-dependent association of THRUMIN1 with 14-3-3 KAPPA and OMEGA were also identified. Together, these studies provide new insights into the mechanistic role of THRUMIN1 in light-dependent chloroplast movements.

scholarly journals Novel anionic cecropins from the spruce budworm feature a poly-L-aspartic acid C-terminus

2018 ◽  
Author(s):  
Halim Maaroufi ◽  
Michel Cusson ◽  
Roger C. Levesque
Keyword(s):  
Anticancer Activity ◽  
Aspartic Acid ◽  
Choristoneura Fumiferana ◽  
Antibacterial Properties ◽  
Spruce Budworm ◽  
Duplication Event ◽  
List Type ◽  
C Terminus ◽  
Genes Encoding ◽  
Dynamics Simulations

AbstractCecropins form a family of amphipathic α-helical cationic peptides with broad-spectrum antibacterial properties and potent anticancer activity. The emergence of bacteria and cancer cells showing resistance to cationic antimicrobial peptides (CAMPs) has fostered a search for new, more selective and more effective alternatives to CAMPs. With this goal in mind, we looked for cecropin homologs in the genome and transcriptome of the spruce budworm, Choristoneura fumiferana. Not only did we find paralogs of the conventional cationic cecropins (Cfcec+), our screening also led to the identification of previously uncharacterized anionic cecropins (Cfcec−), featuring a poly-L-aspartic acid C-terminus. Comparative peptide analysis indicated that the C-terminal helix of Cfcec− is amphipathic, unlike that of Cfcec+, which is hydrophobic. Interestingly, molecular dynamics simulations pointed to the lower conformational flexibility of Cfcec− peptides, relative to that of Cfcec+. Phylogenetic analysis suggests that the evolution of distinct Cfcec+ and Cfcec− peptides may have resulted from an ancient duplication event within the Lepidoptera. Our analyses also indicated that Cfcec− shares characteristics with entericidins, which are involved in bacterial programmed cell death, lunasin, a peptide of plant origins with antimitotic effects, and APC15, a subunit of the anaphase-promoting complex. Finally, we found that both anionic and cationic cecropins contain a BH3-like motif (G-[KQR]-[HKQNR]-[IV]-[KQR]) that could interact with Bcl-2, a protein involved in apoptosis; this observation is congruent with previous reports indicating that cecropins induce apoptosis. Altogether, our observations suggest that cecropins may provide templates for the development of new anticancer drugs.Graphical abstractHighlightsGenes encoding novel anionic cecropins (Cfcec−), featuring a C-terminal poly-L-aspartic acid, were found in the genome of the spruce budworm, Choristoneura fumiferana.Divergence between Cfcec+ and Cfcec− could be the result of an ancient duplication event within the Lepidoptera.There is an apparent relationship between motifs observed in cecropin peptides and apoptosis.Anionic cecropins from the spruce budworm display characteristics suggesting they could have anticancer activity

scholarly journals An Erythrocyte Cytoskeleton-Binding Motif in Exported Plasmodium falciparum Proteins

2011 ◽  
Vol 10 (11) ◽  
pp. 1439-1447 ◽  
Author(s):  
Geoffrey K. Kilili ◽  
Douglas J. LaCount
Keyword(s):  
Plasmodium Falciparum ◽  
Host Cell ◽  
Aspartic Acid ◽  
Binding Motif ◽  
Host Cell Membrane ◽  
Content Type ◽  
C Terminus ◽  
Erythrocyte Surface ◽  
Protein 4.1R ◽  
Erythrocyte Cytoskeleton

ABSTRACTBinding of exported malaria parasite proteins to the host cell membrane and cytoskeleton contributes to the morphological, functional, and antigenic changes seen inPlasmodium falciparum-infected erythrocytes. One such exported protein that targets the erythrocyte cytoskeleton is the mature parasite-infected erythrocyte surface antigen (MESA), which interacts with the N-terminal 30-kDa domain of protein 4.1R via a 19-residue sequence. We report here that the MESA erythrocyte cytoskeleton-binding (MEC) domain is present in at least 13 otherP. falciparumproteins predicted to be exported to the host cell. An alignment of the putative cytoskeleton-binding sequences revealed a conserved aspartic acid at the C terminus that was omitted from the originally reported binding domain. Mutagenesis experiments demonstrated that this aspartic acid was required for the optimal binding of MESA to inside-out vesicles (IOVs) prepared from erythrocytes. Using pulldown assays, we characterized the binding of fragments encoding the MEC domains from PFE0040c/MESA and six other proteins (PF10_0378, PFA0675w, PFB0925w, PFD0095c, PFF1510w, and PFI1790w) to IOVs. All seven proteins bound to IOVs, with MESA showing the strongest affinity in saturation binding experiments. We further examined the interaction of the MEC domain proteins with components of the erythrocyte cytoskeleton and showed that MESA, PF10_0378, and PFA0675w coprecipitated full-length 4.1R from lysates prepared from IOVs. These data demonstrated that the MEC motif is present and functional in at least six otherP. falciparumproteins that are exported to the host cell cytoplasm.

scholarly journals The C-terminus of Wheat streak mosaic virus Coat Protein Is Involved in Differential Infection of Wheat and Maize through Host-Specific Long-Distance Transport

2014 ◽  
Vol 27 (2) ◽  
pp. 150-162 ◽  
Author(s):  
Satyanarayana Tatineni ◽  
Roy French
Keyword(s):  
Amino Acid ◽  
Host Range ◽  
Mosaic Virus ◽  
Aspartic Acid ◽  
Wheat Streak Mosaic Virus ◽  
Coat Proteins ◽  
Long Distance ◽  
Long Distance Transport ◽  
C Terminus ◽  
Distance Transport

Viral determinants and mechanisms involved in extension of host range of monocot-infecting viruses are poorly understood. Viral coat proteins (CP) serve many functions in almost every aspect of the virus life cycle. The role of the C-terminal region of Wheat streak mosaic virus (WSMV) CP in virus biology was examined by mutating six negatively charged aspartic acid residues at positions 216, 289, 290, 326, 333, and 334. All of these amino acid residues are dispensable for virion assembly, and aspartic acid residues at positions 216, 333, and 334 are expendable for normal infection of wheat and maize. However, mutants D289N, D289A, D290A, DD289/290NA, and D326A exhibited slow cell-to-cell movement in wheat, which resulted in delayed onset of systemic infection, followed by a rapid recovery of genomic RNA accumulation and symptom development. Mutants D289N, D289A, and D326A inefficiently infected maize, eliciting milder symptoms, while D290A and DD289/290NA failed to infect systemically, suggesting that the C-terminus of CP is involved in differential infection of wheat and maize. Mutation of aspartic acid residues at amino acid positions 289, 290, and 326 severely debilitated virus ingress into the vascular system of maize but not wheat, suggesting that these amino acids facilitate expansion of WSMV host range through host-specific long-distance transport.

scholarly journals Detection of Isopeptide Bonds in Monoclonal Antibody Aggregates

2021 ◽  
Author(s):  
Thomas Powell ◽  
Michael J. Knight ◽  
Amanda Wood ◽  
John O’Hara ◽  
William Burkitt
Keyword(s):  
Monoclonal Antibody ◽  
Aspartic Acid ◽  
Peptide Mapping ◽  
Chemical Mechanism ◽  
Size Exclusion ◽  
C Terminus ◽  
Lysine Residues ◽  
Aggregate Fractions ◽  
Isopeptide Bonds ◽  
First Time

Abstract Purpose A major difficulty in monoclonal antibody (mAb) therapeutic development is product aggregation. In this study, intermolecular isopeptide bonds in mAb aggregates were characterized for the first time. We aim to propose a mechanism of covalent aggregation in a model antibody using stressed studies at raised temperatures to aid in the understanding of mAb aggregation pathways. Methods Aggregate fractions were generated using raised temperature and were purified using size-exclusion chromatography (SEC). The fractions were tryptically digested and characterized using liquid chromatography hyphenated to tandem mass-spectrometry (LC–MS/MS). Results An increased amount of clipping between aspartic acid and proline in a solvent accessible loop in the constant heavy 2 (CH2) domain of the mAb was observed under these conditions. Detailed peptide mapping revealed 14 isopeptide bonds between aspartic acid at that cleavage site and lysine residues on adjacent antibodies. Two additional isopeptide bonds were identified between the mAb HC N-terminal glutamic acid or a separate aspartic acid to lysine residues on adjacent antibodies. Conclusions Inter-protein isopeptide bonds between the side chains of acidic amino acids (aspartate and glutamate) and lysine were characterized for the first time in mAb aggregates. A chemical mechanism was presented whereby spontaneous isopeptide bond formation could be facilitated via either the aspartic acid side chain or C-terminus.

scholarly journals The Retrieval Function of the KDEL Receptor Requires PKA Phosphorylation of Its C-Terminus

2003 ◽  
Vol 14 (10) ◽  
pp. 4114-4125 ◽  
Author(s):  
Margarita Cabrera ◽  
Manuel Muñiz ◽  
Josefina Hidalgo ◽  
Lucia Vega ◽  
María Esther Martín ◽  
...  
Keyword(s):  
Aspartic Acid ◽  
Retrograde Transport ◽  
Integral Membrane Protein ◽  
Truncated Form ◽  
Permeabilized Cells ◽  
C Terminus ◽  
Intermediate Compartment ◽  
Peptide Binding Assay ◽  
Kdel Receptor

The KDEL receptor is a Golgi/intermediate compartment-located integral membrane protein that carries out the retrieval of escaped ER proteins bearing a C-terminal KDEL sequence. This occurs throughout retrograde traffic mediated by COPI-coated transport carriers. The role of the C-terminal cytoplasmic domain of the KDEL receptor in this process has been investigated. Deletion of this domain did not affect receptor subcellular localization although cells expressing this truncated form of the receptor failed to retain KDEL ligands intracellularly. Permeabilized cells incubated with ATP and GTP exhibited tubular processes-mediated redistribution from the Golgi area to the ER of the wild-type receptor, whereas the truncated form lacking the C-terminal domain remained concentrated in the Golgi. As revealed with a peptide-binding assay, this domain did not interact with both coatomer and ARF-GAP unless serine 209 was mutated to aspartic acid. In contrast, alanine replacement of serine 209 inhibited coatomer/ARF-GAP recruitment, receptor redistribution into the ER, and intracellular retention of KDEL ligands. Serine 209 was phosphorylated by both cytosolic and recombinant protein kinase A (PKA) catalytic subunit. Inhibition of endogenous PKA activity with H89 blocked Golgi-ER transport of the native receptor but did not affect redistribution to the ER of a mutated form bearing aspartic acid at position 209. We conclude that PKA phosphorylation of serine 209 is required for the retrograde transport of the KDEL receptor from the Golgi complex to the ER from which the retrieval of proteins bearing the KDEL signal depends.

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