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Comparison of Substrate Specificity of <em>escherichia Coli</em>p-Aminobenzoyl-Glutamate Hy-drolase with <em>Pseudomonas</em>Carboxypeptidase G

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Advances in enzyme Research 2014年第1期2卷 39-48页

作者： cassandra M. Larimer Dejan Slavnic Lenore D. Pitstick Jacalyn M. Green Department of Biochemistry Chicago College of Osteopathic Medicine Midwestern University Downers Grove IL USA

Reduced folic acid derivatives support biosynthesis of DNA, RNA and amino acids in bacteria as well as in eukaryotes, including humans. While the genes and steps for bacterial folic acid synthesis are known, those associated with folic acid catabolism are not well understood. A folate catabolite found in both humans and bacteria is p-aminobenzoyl-glutamate (PABA-GLU). The enzyme p-aminobenzoyl-glutamate hydrolase (PGH) breaks down PABA-GLU and is part of an apparent operon, the abg region, in e. coli. The subunits of PGH possess sequence and catalytic similarities to carboxypeptidase enzymes from Pseudomonas species. A comparison of the subunit sequences and activity of PGH, relative to carboxypeptidase enzymes, may lead to a better understanding of bacterial physiology and pathway evolution. We first compared the amino acid sequences of AbgA, AbgB and carboxypeptidase G2 from Pseudomonas sp. RS-16, which has been crystallized. Then we compared the enzyme activities of e. coli PGH and commercially available Pseudomonas carboxypeptidase G using spectrophotometric assays measuring cleavage of PABA-GLU, folate, aminopterin, methotrexate, 5-formyltetrahydrofolate, and 5-methyltetrahydrofolate. The Km and Vmax values for the folate and anti-folate substrates of PGH could not be determined, because the instrument reached its limit before the enzyme was saturated. Therefore, activity of PGH was compared to the activity of CPG, or normalized to PABA-GLU (nmole/min/μg). Relative to its activity with 10 μM PABA-GLU (100%), PGH cleaved glutamate from methotrexate (48%), aminopterin (45%) and folate (9%). Reduced folates leucovorin (5-formyltetrahydrofolate) and 5-methyltetrahydrofolate were not cleaved by PGH. Our data suggest that e. coli PGH is specific for PABA-GLU as its activity with natural folates (folate, 5-methyltetrahydrofolate, and leucovorin) was very poor. It does, however, have some ability to cleave anti-folates which may have clinical applications in treatment

关键词： Folate p-Aminobenzoyl-Glutamate Hydrolase Carboxypeptidase G Folate Catabolism

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SARS-CoV-2 productively infects primary human immune system cells in vitro and in CoviD-19 patients

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Journal of Molecular Cell Biology 2022年第4期14卷 51-64页

作者： Marjorie C.Pontelli italo A.Castro Ronaldo B.Martins Leonardo La Serra Flavio P.Veras Daniele C.Nascimento Camila M.Silva Ricardo S.Cardoso Roberta Rosales Rogério Gomes Thais M.Lima Juliano P.Souza Brenda C.Vitti Diego B.Caetite Mikhael H.F.de Lima Spencer D.Stumpf cassandra e.Thompson Louis-Marie Bloyet Juliana e Toller-Kawahisa Marcela C.Giannini Leticia P.Bonjorno Marial.F.Lopes Sabrina S.Batah Li Siyuan Rodrigo Luppino-Assad Sergio C.L.Almeida Fabiola R.Oliveira Maira N.Benatti Lorena L.F.Pontes Rodrigo C.Santana Fernando C.Vilar Maria Auxiliadora-Martins Pei-Yong Shi Thiago M.Cunha Rodrigo T.Calado JoséC.Alves-Filho Dario S.Zamboni Alexandre T.Fabro Paulo Louzada-Junior Rene D.R.Oliveira Sean P.J.Whelan Fernando Q.Cunha eurico Arruda Virology Research Center Ribeirao Preto Medical SchoolUniversity of Sao Pauloibeirao PretoSao Paulo 14049-900.Brazil Center of Research in Inflammatory Diseases Ribeirao Preto Medical SchoolUniversity of Sao PauloRibeirao PretoSao Paulo 14049-900Brazil Department of Celland Molecular Biology Ribeirao Preto Medical SchoolUniversityof Sao PauloRibeirao PretoSao Paulo 14049-900 Brazil Regional Blood Centerof Ribeirao Preto Ribeirao Preto Medical ScholUniversity of Sao PauloRibeirao PretoSao Paulo 14049-900Brazil Department of Biochemistry&Molecular Biology the University of Texas Medical BranchGalvestonTX7755USA Divisions of Clinical lmmunology Infectious Diseases and Intensive Care UnitRibeirao Preto Medical Schooniversity of Sao PauloRibeirao PretoSao Paulo 14049-900Brazil Department of Pathology Ribeirao Preto Medical Scholniversity of Sao PauloRibeirao PretoSao Paulo 14049-900Brazil Department of Molecular Microbiology.Washington University in St.Louis Saint LouisMO 63110USA

The severe acute respiratory syndrome coronavirus 2(SARS-CovV-2)infection is associated with a hyperinflammatory state and lymphocytopenia,a hallmark that appears as both signature and prognosis of disease severity *** cytokine storm and a sustained inflammatory state are commonly associated with immune cell depletion,it is still unclear whether direct SARS-Cov-2 infection of immune cells could also play a role in this scenario by harboring viral *** found that monocytes,as well as both B and T lymphocytes,were susceptible to SARS-Cov-2 infection in vitro,accumulating double-stranded RNA consistent with viral RNA replication and ultimately leading to expressive T cell *** addition,flow cytometry and immunofluorescence analysis revealed that SARS-Cov-2 was frequently detected in monocytes and B lymphocytes from coronavirus disease 2019(CoVID-19)*** rates of SARS-Cov-2-infected monocytes in peripheral blood mononuclear cells from CoVID-19 patients increased over time from symptom onset,with SARS-CoV-2-positive monocytes,B cells,and CD4+T lymphocytes also detected in postmortem lung *** results indicated that SARS-CoV-2 infection of blood-circulating leukocytes in covID-19 patients might have important implications for disease pathogenesis and progression,immune dysfunction,and virus spread within the host.

关键词： SARS-CoV-2 CoVID-19 lymphocytopenia peripheral blood mononuclear cell(PBMC) lymphocytes monocytes apoptosis

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Semi-Automated Method for Multi-Tasking Measurement of Microbial Growth, Capsule, and Biofilm Formation

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Advances in Microbiology 2012年第4期2卷 623-628页

作者： Derek T. Kirby Craigory J. Raino Samuel F. Rabor Jr. cassandra J. Wasson Balbina J. Plotkin Department of Microbiology and Immunology Midwestern University Downers Grove USA Department of Microbiology and Immunology Midwestern University Downers Grove USA

The majority of bacterial infections involve the formation of biofilms. Biofilm formation is nutrient and growth dependent. Determination of the effects of nutrients on exopolysaccharide production and bacterial growth is labor and time intensive. We tested whether the Bioscreen C (Growth Curves, Inc.) would have utility as a high-throughput tool in the measurement of fundamental phenotype expression, as it relates to growth conditions. Within 48 - 72 hr, reproduceble, statistically significant data on the affects of growth conditions on generation time, capsule production and biofilm formation (maximally for 25 different conditions per 24 hr run cycle;n = 4) were obtained. Although all S. aureus strains produced similar amounts of capsule, sarA– and agr– strains grew significantly slower than parent strain (1.6 fold slower) and produced significantly (p 0.05) less biofilm (~2 fold). e. coli growth rate, biofilm and capsule production in simulated nephropathic urine medium was similar for urine with insulin (20 μU). Addition of insulin to urine medium with proline increased generation time, capsule and biofilm production. Findings from this study show that the Bioscreen C is a rapid, reproducible, and easily manipulated system to concurrently measure bacterial growth, biofilm formation, and capsule production. In addition, there is the potential for further applications of this system by expanding the types of detector dyes used.

关键词： Biofilm Staphylococcus aureus escherichia coli Quorum-Signaling Compounds Menaquinone Insulin Growth

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Activation of RAS/MAPK pathway confers MCL-1 mediated acquired resistance to BCL-2 inhibitor venetoclax in acute myeloid leukemia

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Signal Transduction and Targeted Therapy 2022年第3期7卷 856-868页

作者： Qi Zhang Bridget Riley-Gillis Lina Han Yanan Jia Alessia Lodi Haijiao Zhang Saravanan Ganesan Rongqing Pan Sergej N.Konoplev Shannon R.Sweeney Jeremy A.Ryan Yulia Jitkova Kenneth Dunner Jr Shaun e.Grosskurth Priyanka Vijay Sujana Ghosh Charles Lu Wencai Ma Stephen Kurtz Vivian R.Ruvolo Helen Ma Connie CWeng cassandra LRamage Natalia Baran Ce Shi Tianyu Cai Richard eric Davis Venkata L.Battula Yingchang Mi Jing Wang Courtney D.DiNardo Michael Andreeff Jeffery W.Tyner Aaron Schimmer Anthony Letai Rose Ann Padua Carlos e.Bueso-Ramos Stefano Tiziani Joel Leverson Relja Popovic Marina Konopleva Department of Leukemia The University of Texas MD Anderson Cancer CenterHoustonTXUSA AbbVie Inc. North ChicagoILUSA Institute of Hematology Chinese Academy of Medical Sciences&Peking Union Medical CollegeTianjinChina Department of Nutritional Sciences Department of PediatricsDepartment of OncologyDell Medical SchoolThe University of Texas at AustinAustinTX 78712USA Department of Cell Developmental&Cancer BiologyDivision of Hematology&Medical OncologyKnight Cancer InstituteOregon Health&Science UniversityPortlandORUSA Universite de Paris Institut de la Recherche Saint-Louis(IRSL)Inserm Unit 1131ParisFrance Dana-Farber Cancer Institute BostonMAUSA Department of Hematopathology The University of Texas MD Anderson Cancer CenterHoustonTXUSA Princess Margaret Cancer Center TorontoONCanada High Resolution electron Microscopy Facility The University of Texas MD Anderson Cancer CenterHoustonTXUSA Department of Bioinformatics&Computational Biology The University of Texas MD Anderson Cancer CenterHoustonTXUSA Department of Hematology The First Hospital Affiliated Harbin Medical UniversityHarbinChina Department of Lymphoma&Myeloma Research The University of Texas MD Anderson Cancer CenterHoustonTXUSA

Despite high initial response rates,acute myeloid leukemia(AML)treated with the BCL-2-selective inhibitor venetoclax(VeN)alone or in combinations commonly acquires *** performed gene/protein expression,metabolomic and methylation analyses of isogenic AML cell lines sensitive or resistant to VeN,and identified the activation of RAS/MAPK pathway,leading to increased stability and higher levels of MCL-1 protein,as a major acquired mechanism of VeN resistance.

关键词： acute myeloid acquired

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Correction: Activation of RAS/MAPK pathway confers MCL-1 mediated acquired resistance to BCL-2 inhibitor venetoclax in acute myeloid leukemia

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Signal Transduction and Targeted Therapy 2022年第5期7卷 1860-1860页

作者： Qi Zhang Bridget Riley-Gillis Lina Han Yannan Jia Alessia Lodi Haijiao Zhang Saravanan Ganesan Rongqing Pan Sergej N.Konoplev Shannon R.Sweeney Jeremy A.Ryan Yulia Jitkova Kenneth Dunner Jr Shaun e.Grosskurth Priyanka Vijay Sujana Ghosh Charles Lu Wencai Ma Stephen Kurtz Vivian R.Ruvolo Helen Ma Connie C.Weng cassandra L.Ramage Natalia Baran Ce Shi Tianyu Cai Richard eric Davis Venkata L.Battula Yingchang Mi Jing Wang Courtney D.DiNardo Michael Andreeff Jeffery W.Tyner Aaron Schimmer Anthony Letai Rose Ann Padua Carlos e.Bueso-Ramos Stefano Tiziani Joel Leverson Relja Popovic Marina Konopleva Department of Leukemia The University of Texas MD Anderson Cancer Center Houston TX USA Institute of Hematology Chinese Academy of Medical Sciences & Peking Union Medical College Tianjin China Department of Hematology The First Hospital Affiliated Harbin Medical University Harbin China AbbVie Inc. North Chicago IL USA Department of Nutritional Sciences Department of Pediatrics Department of Oncology Dell Medical School The University of Texas at Austin Austin TX 78712 USA Department of Cell Developmental & Cancer Biology Division of Hematology & Medical Oncology Knight Cancer Institute Oregon Health & Science University Portland OR USA Université de Paris Institut de la Recherche Saint-Louis (IRSL) Inserm Unit 1131 Paris France Dana-Farber Cancer Institute Boston MA USA Department of Hematopathology The University of Texas MD Anderson Cancer Center Houston TX USA Princess Margaret Cancer Center Toronto ON Canada High Resolution electron Microscopy Facility The University of Texas MD Anderson Cancer Center Houston TX USA Department of Bioinformatics & Computational Biology The University of Texas MD Anderson Cancer Center Houston TX USA Department of Lymphoma & Myeloma Research The University of Texas MD Anderson Cancer Center Houston TX USA

Correction to:Signal Transduction and Targeted Therapy(2022)7:1–13,https://***/10.1038/s41392-021-00870-3 In the original version of this article,given name of 4th author Yannan Jia was incorrectly published as Yanan *** original article has been *** Access This article is licensed under a Creative Commons Attribution 4.0 International License,which permits use,sharing,adaptation,distribution and reproduction in any medium or format,as long as you give appropriate credit to the original author(s)and the source.

关键词： myeloid Therapy resistance

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