scholarly journals Infrequent Occurrence of Natural Mutations in the pp65495–503 Epitope Sequence Presented by the HLA A∗0201 Allele among Human Cytomegalovirus Isolates

2001 ◽  
Vol 75 (5) ◽  
pp. 2472-2474 ◽  
Author(s):  
John A. Zaia ◽  
Ghislaine Gallez-Hawkins ◽  
Xiuli Li ◽  
Zhi-Qiang Yao ◽  
Norma Lomeli ◽  
...  
Keyword(s):  
Human Cytomegalovirus ◽  
Peptide Sequence ◽  
Amino Terminus ◽  
Methionine Residue ◽  
Rare Mutations ◽  
Epitope Sequence ◽  
Human Cmv ◽  
Specific Peptide

ABSTRACT To determine if mutations of an immunodominant HLA-restricted cytomegalovirus (CMV) peptide sequence occur in nature, the sequence corresponding to the HLA A∗0201-specific peptide CMVpp65495–503 was determined in 50 human CMV isolates. Rare mutations were detected; 6 of 50 were silent mutations at the amino terminus of the peptide, while 3 of 50 were mutations of the native methionine residue to isoleucine (M499I). The observed M499I mutation in three isolates decreased cytolytic targeting.

scholarly journals The Chlamydomonas Dhc1 gene encodes a dynein heavy chain subunit required for assembly of the I1 inner arm complex.

1997 ◽  
Vol 8 (4) ◽  
pp. 607-620 ◽  
Author(s):  
S H Myster ◽  
J A Knott ◽  
E O'Toole ◽  
M E Porter
Keyword(s):  
Heavy Chain ◽  
Insertional Mutagenesis ◽  
Northern Blot Analysis ◽  
Immunoblot Analysis ◽  
Transcription Unit ◽  
Peptide Sequence ◽  
Dynein Heavy Chain ◽  
The Individual ◽  
Intermediate Chains ◽  
Specific Peptide

Multiple members of the dynein heavy chain (Dhc) gene family have been recovered in several organisms, but the relationships between these sequences and the Dhc isoforms that they encode are largely unknown. To identify Dhc loci and determine the specific functions of the individual Dhc isoforms, we have screened a collection of motility mutants generated by insertional mutagenesis in Chlamydomonas. In this report, we characterize one strain, pf9-3, in which the insertion event was accompanied by a deletion of approximately 13 kb of genomic DNA within the transcription unit of the Dhc1 gene. Northern blot analysis confirms that pf9-3 is a null mutation. Biochemical and structural studies of isolated axonemes demonstrate that the pf9-3 mutant fails to assemble the I1 inner arm complex, a two-headed dynein isoform composed of two Dhcs (1 alpha and 1 beta) and three intermediate chains. To determine if the Dhc1 gene product corresponds to one of the Dhcs of the I1 complex, antibodies were generated against a Dhc1-specific peptide sequence. Immunoblot analysis reveals that the Dhc1 gene encodes the 1 alpha Dhc subunit. These studies thus, identify the first inner arm Dhc locus to be described in any organism and further demonstrate that the 1 alpha Dhc subunit plays an essential role in the assembly of the I1 inner arm complex.

Identification of ADAMTS13 Peptide Sequences Recognized by Autoantibodies in Patients with Acquired Thrombotic Thrombocytopenic Purpura.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2286-2286
Author(s):  
Yusuke Yamaguchi ◽  
Takanori Moriki ◽  
Hideo Wada ◽  
Masanori Matsumoto ◽  
Yoshihiro Fujimura ◽  
...  
Keyword(s):  
Thrombotic Thrombocytopenic Purpura ◽  
Peptide Sequence ◽  
Adamts13 Activity ◽  
Lambda Phage ◽  
Thrombocytopenic Purpura ◽  
C Terminus ◽  
Epitope Sequence ◽  
Von Willebrand ◽  
The Impact ◽  
Inhibitor Titer

Abstract Anti-ADAMTS13 autoantibodies are considered to play pivotal roles in the pathophysiology of acquired thrombotic thrombocytopenic purpura (TTP). They inhibit the ADAMTS13 function resulting in the appearance of ultra-large von Willebrand factor (VWF) multimers. Major binding sites of the autoantibodies were reported to be in the cysteine-rich/spacer domains. To clarify the precise peptide sequences recognized by anti-ADAMTS13 IgG autoantibodies, we constructed a random cDNA fragment library expressing various peptides of ADAMTS13 on the surface of lambda phage and screened the library using purified IgG from 13 TTP patients. Diverse peptide sequences were obtained from almost entire ADAMTS13 domains such as metalloprotease, disintegrin, TSP1-1, cysteine-rich, spacer, TSP1- 2, 3, 4, 5, 7, 8 and CUB1. In particular, we detected an identical 26 amino-acid epitope sequence in the C-terminus of spacer domain from Gly662 to Val687 (sp662–687) shared by 5 TTP patients. Moreover, the peptide sequence was exactly included in one of the VWF binding epitope sites that we previously determined (Blood110 (11), 795a, 2007). We then assessed the impact of specific autoantibody to ADAMTS13 activity measured by FRETS-VWF73 or EIA and ADAMTS13 inhibitor titer in each of TTP patient plasma. However, both of the ADAMTS13 activity and inhibitor titer seemed not correlated with the existence of specific sp662–687 IgG autoantibody. These observations suggest that the autoantibody to sp662–687 may be one specific feature of TTP, although other epitopes are also involved in the pathogenesis of the disorder.

scholarly journals Breast Milk Human Cytomegalovirus (CMV) Viral Load and the Establishment of Breast Milk CMV-pp65-Specific CD8 T Cells in Human CMV Infected Mothers

2017 ◽  
Vol 216 (9) ◽  
pp. 1176-1179 ◽  
Author(s):  
David C Moylan ◽  
Sunil K Pati ◽  
Shannon A Ross ◽  
Karen B Fowler ◽  
Suresh B Boppana ◽  
...  
Keyword(s):  
T Cells ◽  
Viral Load ◽  
Breast Milk ◽  
Human Cytomegalovirus ◽  
Cd8 T Cells ◽  
Human Cmv

scholarly journals A Mutation Deleting Sequences Encoding the Amino Terminus of Human Cytomegalovirus UL84 Impairs Interaction with UL44 and Capsid Localization

2012 ◽  
Vol 86 (20) ◽  
pp. 11066-11077 ◽  
Author(s):  
B. L. Strang ◽  
B. J. Bender ◽  
M. Sharma ◽  
J. M. Pesola ◽  
R. L. Sanders ◽  
...  
Keyword(s):  
Human Cytomegalovirus ◽  
Amino Terminus

scholarly journals A Combination Strategy of Solubility Enhancers for Effective Production of Soluble and Bioactive Human Enterokinase

2021 ◽  
Author(s):  
Jinhak Kwon ◽  
Hyeongjun Cho ◽  
Seungmin Kim ◽  
Yiseul Ryu ◽  
Joong-jae Lee
Keyword(s):  
Recombinant Proteins ◽  
Phosphoglycerate Kinase ◽  
Combinatorial Design ◽  
Peptide Sequence ◽  
Protein Digestion ◽  
Combination Strategy ◽  
Enhanced Solubility ◽  
Production Strategy ◽  
Solubility Enhancers ◽  
Specific Peptide

Enterokinase is one of the hydrolases that catalyze hydrolysis to regulate biological processes in intestinal visceral mucosa. Enterokinase plays an essential role in accelerating the process of protein digestion as it converts trypsinogen into active trypsin by accurately recognizing and cleaving a specific peptide sequence, (Asp)4-Lys. Due to its exceptional substrate specificity, enterokinase is widely used as a versatile molecular tool in various bioprocessing, especially in removing fusion tags from recombinant proteins. Despite its biotechnological importance, mass production of soluble enterokinase in bacteria still remains an unsolved challenge. Here, we present an effective production strategy of human enterokinase using tandemly linked solubility enhancers consisting of thioredoxin, phosphoglycerate kinase or maltose-binding protein. The resulting enterokinases exhibited significantly enhanced solubility and bacterial expression level while retaining enzymatic activity, which demonstrates that combinatorial design of fusion proteins has the potential to provide an efficient way to produce recombinant proteins in bacteria.

scholarly journals Human Cytomegalovirus US2 Endoplasmic Reticulum-Lumenal Domain Dictates Association with Major Histocompatibility Complex Class I in a Locus-Specific Manner

2001 ◽  
Vol 75 (11) ◽  
pp. 5197-5204 ◽  
Author(s):  
Benjamin E. Gewurz ◽  
Evelyn W. Wang ◽  
Domenico Tortorella ◽  
Danny J. Schust ◽  
Hidde L. Ploegh
Keyword(s):  
Endoplasmic Reticulum ◽  
Major Histocompatibility Complex ◽  
Mhc Class I ◽  
Human Cytomegalovirus ◽  
Peptide Sequence ◽  
Class I ◽  
Major Histocompatibility ◽  
Independent Manner ◽  
Histocompatibility Complex ◽  
Lumenal Domain

ABSTRACT The human cytomegalovirus-encoded US2 glycoprotein targets endoplasmic reticulum-resident major histocompatibility complex (MHC) class I heavy chains for rapid degradation by the proteasome. We demonstrate that the endoplasmic reticulum-lumenal domain of US2 allows tight interaction with class I molecules encoded by the HLA-A locus. Recombinant soluble US2 binds properly folded, peptide-containing recombinant HLA-A2 molecules in a peptide sequence-independent manner, consistent with US2's ability to broadly downregulate class I molecules. The physicochemical properties of the US2/MHC class I complex suggest a 1:1 stoichiometry. These results demonstrate that US2 does not require additional cellular proteins to specifically interact with soluble class I molecules. Binding of US2 does not significantly alter the conformation of class I molecules, as a soluble T-cell receptor can simultaneously recognize class I molecules associated with US2. The lumenal domain of US2 can differentiate between the products of distinct class I loci, as US2 binds several HLA-A locus products while being unable to bind recombinant HLA-B7, HLA-B27, HLA-Cw4, or HLA-E. We did not observe interaction between soluble US2 and either recombinant HLA-DR1 or recombinant HLA-DM. The substrate specificity of US2 may help explain the presence in human cytomegalovirus of multiple strategies for downregulation of MHC class I molecules.

scholarly journals Coiled-Coil Domains in Glycoproteins B and H Are Involved in Human Cytomegalovirus Membrane Fusion

2004 ◽  
Vol 78 (15) ◽  
pp. 8333-8341 ◽  
Author(s):  
Matthew Lopper ◽  
Teresa Compton
Keyword(s):  
Membrane Fusion ◽  
Human Cytomegalovirus ◽  
Virus Entry ◽  
Coiled Coil ◽  
Coiled Coils ◽  
Viral Envelope ◽  
Heptad Repeat ◽  
Repeat Sequences ◽  
Human Cmv ◽  
Hsv 1

ABSTRACT Human cytomegalovirus (CMV) utilizes a complex route of entry into cells that involves multiple interactions between viral envelope proteins and cellular receptors. Three conserved viral glycoproteins, gB, gH, and gL, are required for CMV-mediated membrane fusion, but little is known of how these proteins cooperate during entry (E. R. Kinzler and T. Compton, submitted for publication). The goal of this study was to begin defining the molecular mechanisms that underlie membrane fusion mediated by herpesviruses. We identified heptad repeat sequences predicted to form alpha-helical coiled coils in two glycoproteins required for fusion, gB and gH. Peptides derived from gB and gH containing the heptad repeat sequences inhibited virus entry when introduced coincident with virus inoculation onto cells or when mixed with virus prior to inoculation. Neither peptide affected binding of CMV to fibroblasts, suggesting that the peptides inhibit membrane fusion. Both gB and gH coiled-coil peptides blocked entry of several laboratory-adapted and clinical strains of human CMV, but neither peptide affected entry of murine CMV or herpes simplex virus type 1 (HSV-1). Although murine CMV and HSV-1 gB and gH have heptad repeat regions, the ability of human CMV gB and gH peptides to inhibit virus entry correlates with the specific residues that comprise the heptad repeat region. The ability of gB and gH coiled-coil peptides to inhibit virus entry independently of cell contact suggests that the coiled-coil regions of gB and gH function differently from those of class I, single-component fusion proteins. Taken together, these data support a critical role for alpha-helical coiled coils in gB and gH in the entry pathway of CMV.

scholarly journals A predictive model for vertebrate bone identification from collagen using proteomic mass spectrometry

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Heyi Yang ◽  
Erin R. Butler ◽  
Samantha A. Monier ◽  
Jennifer Teubl ◽  
David Fenyö ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Species Identification ◽  
Probabilistic Approach ◽  
Modern Human ◽  
Peptide Sequence ◽  
Human Species ◽  
Interpretation Errors ◽  
Species Specific ◽  
Sequence Errors ◽  
Specific Peptide

AbstractProteogenomics is an increasingly common method for species identification as it allows for rapid and inexpensive interrogation of an unknown organism’s proteome—even when the proteome is partially degraded. The proteomic method typically uses tandem mass spectrometry to survey all peptides detectable in a sample that frequently contains hundreds or thousands of proteins. Species identification is based on detection of a small numbers of species-specific peptides. Genetic analysis of proteins by mass spectrometry, however, is a developing field, and the bone proteome, typically consisting of only two proteins, pushes the limits of this technology. Nearly 20% of highly confident spectra from modern human bone samples identify non-human species when searched against a vertebrate database—as would be necessary with a fragment of unknown bone. These non-human peptides are often the result of current limitations in mass spectrometry or algorithm interpretation errors. Consequently, it is difficult to know if a “species-specific” peptide used to identify a sample is actually present in that sample. Here we evaluate the causes of peptide sequence errors and propose an unbiased, probabilistic approach to determine the likelihood that a species is correctly identified from bone without relying on species-specific peptides.

scholarly journals MODERN ETHIOTROPIC CHEMOTHERAPY OF HUMAN CYTOMEGALOVIRUS INFECTION: CLINICAL EFFECTIVENESS, MOLECULAR MECHANISM OF ACTION, DRUG RESISTANCE, NEW TRENDS AND PROSPECTS. PART 2

2018 ◽  
Vol 63 (6) ◽  
pp. 250-260
Author(s):  
V. L. Andronova
Keyword(s):  
Drug Resistance ◽  
Human Cytomegalovirus ◽  
Clinical Effectiveness ◽  
Great Activity ◽  
Third Phase ◽  
Viral Cyclin ◽  
Preclinical Trials ◽  
Molecular Mechanism Of Action ◽  
Resistant Strains ◽  
Human Cmv

A number of synthetic compounds, such as the nucleoside analog ganciclovir, its L-valine ester (a metabolic precursor of ganciclovir) and pyrophosphate analog foscarnet, are permitted for the treatment of HCMV-related diseases in the WHO European Region. The viral DNA- polymerase is used by all these drugs as a bio-target. However, the usage of standard anti-CMV therapy is accompanied by severe side effects, as well as the development of drug resistance in the virus, mainly in conditions of immunodeficiency. In this review, we focused on viral proteins of interest as new potential targets and their inhibitors, such as the inhibitor of human CMV terminology, lethermovir, which showed great activity in the third phase of clinical trials, inhibitors of viral cyclin-dependent kinase (maribavir, cyclopropavir ) and a number of compounds exhibiting anti-HCMV-activity, undergoing only preclinical trials in the experiment. Inclusion of new anti-CMV agents that are active against GСV/PFA/CDV-resistant strains of CMV into standard prophylactic and therapeutic regimens, will allow to increase the effectiveness of anti-CMV therapy, including in cases when standard therapy is ineffective. Areas covered: the international databases such as A MEDLINE, PubMed, eLIBRARY.RU, ClinicalTrials.gov., etc. with the purpose of obtaining information on compounds showing selective action against the human cytomegalovirus, the most promising for the development of drugs.

Sign in / Sign up

Export Citation Format

Share Document