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Article ; Online: Detection of Microcrystals for CryoEM.

Weiss, Simon / Vergara, Sandra / Lin, Guowu / Calero, Guillermo

Methods in molecular biology (Clifton, N.J.)

2020 Volume 2215, Page(s) 299–307

Abstract: Here, we present a strategy to identify microcrystals from initial protein crystallization screen experiments and to optimize diffraction quality of those crystals using negative stain transmission electron microscopy (TEM) as a guiding technique. The ... ...

Abstract	Here, we present a strategy to identify microcrystals from initial protein crystallization screen experiments and to optimize diffraction quality of those crystals using negative stain transmission electron microscopy (TEM) as a guiding technique. The use of negative stain TEM allows visualization along the process and thus enables optimization of crystal diffraction by monitoring the lattice quality of crystallization conditions. Nanocrystals bearing perfect lattices are seeded and can be used for MicroED as well as growing larger crystals for X-ray and free electron laser (FEL) data collection.
MeSH term(s)	Cryoelectron Microscopy/methods ; Crystallization/methods ; Microscopy, Electron, Transmission/methods ; Nanoparticles/chemistry ; Nanoparticles/ultrastructure ; Protein Conformation
Language	English
Publishing date	2020-11-09
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-0966-8_14
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Article: Preclinical assessment of a novel human antibody VH domain targeting mesothelin as an antibody-drug conjugate.

Sun, Zehua / Chu, Xiaojie / Adams, Cynthia / Ilina, Tatiana V / Guerrero, Michel / Lin, Guowu / Chen, Chuan / Jelev, Dontcho / Ishima, Rieko / Li, Wei / Mellors, John W / Calero, Guillermo / Dimitrov, Dimiter S

Molecular therapy oncolytics

2023 Volume 31, Page(s) 100726

Abstract: Mesothelin (MSLN) has been a validated tumor-associated antigen target for several solid tumors for over a decade, making it an attractive option for therapeutic interventions. Novel antibodies with high affinity and better therapeutic properties are ... ...

Abstract	Mesothelin (MSLN) has been a validated tumor-associated antigen target for several solid tumors for over a decade, making it an attractive option for therapeutic interventions. Novel antibodies with high affinity and better therapeutic properties are needed. In the current study, we have isolated and characterized a novel heavy chain variable (VH) domain 3C9 from a large-size human immunoglobulin VH domain library. 3C9 exhibited high affinity (KD [dissociation constant] <3 nM) and binding specificity in a membrane proteome array (MPA). In a mouse xenograft model, 3C9 fused to human IgG1 Fc was detected at tumor sites as early as 8 h post-infusion and remained at the site for over 10 days. Furthermore, 3C9 fused to a human Fc domain drug conjugate effectively inhibited MSLN-positive tumor growth in a mouse xenograft model. The X-ray crystal structure of full-length MSLN in complex with 3C9 reveals interaction of the 3C9 domains with two distinctive residue patches on the MSLN surface. This newly discovered VH antibody domain has a high potential as a therapeutic candidate for MSLN-expressing cancers.
Language	English
Publishing date	2023-09-13
Publishing country	United States
Document type	Journal Article
ISSN	2372-7705
ISSN	2372-7705
DOI	10.1016/j.omto.2023.09.002
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Article: Structural basis of transcription: RNA Polymerase II substrate binding and metal coordination at 3.0 Å using a free-electron laser.

Lin, Guowu / Barnes, Christopher O / Weiss, Simon / Dutagaci, Bercem / Qiu, Chenxi / Feig, Michael / Song, Jihnu / Lyubimov, Artem / Cohen, Aina E / Kaplan, Craig D / Calero, Guillermo

bioRxiv : the preprint server for biology

2023

Abstract: Catalysis and translocation of multi-subunit DNA-directed RNA polymerases underlie all cellular mRNA synthesis. RNA polymerase II (Pol II) synthesizes eukaryotic pre-mRNAs from a DNA template strand buried in its active site. Structural details of ... ...

Abstract	Catalysis and translocation of multi-subunit DNA-directed RNA polymerases underlie all cellular mRNA synthesis. RNA polymerase II (Pol II) synthesizes eukaryotic pre-mRNAs from a DNA template strand buried in its active site. Structural details of catalysis at near atomic resolution and precise arrangement of key active site components have been elusive. Here we present the free electron laser (FEL) structure of a matched ATP-bound Pol II, revealing the full active site interaction network at the highest resolution to date, including the trigger loop (TL) in the closed conformation, bonafide occupancy of both site A and B Mg
Language	English
Publishing date	2023-09-22
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.09.22.559052
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Article: Transcription with a laser: Radiation-damage-free diffraction of RNA Polymerase II crystals

Lin, Guowu / Weiss, Simon C / Vergara, Sandra / Camacho, Carlos / Calero, Guillermo

Methods. 2019 Apr. 15, v. 159-160

2019

Abstract: Well-diffracting crystals are essential to obtain relevant structural data that will lead to understanding of RNA Polymerase II (Pol II) transcriptional processes at a molecular level. Here we present a strategy to study Pol II crystals using negative ... ...

Abstract	Well-diffracting crystals are essential to obtain relevant structural data that will lead to understanding of RNA Polymerase II (Pol II) transcriptional processes at a molecular level. Here we present a strategy to study Pol II crystals using negative stain transmission electron microscopy (TEM) and a methodology to optimize radiation damage free data collection using free electron laser (FEL) at the Linac Coherent Light Source (LCLS). The use of negative stain TEM allowed visualization and optimization of crystal diffraction by monitoring the lattice quality of crystallization conditions. Nano crystals bearing perfect lattices were seeded and used to grow larger crystals for FEL data collection. Moreover, the use of in house designed crystal loops together with ultra-violet (UV) microscopy for crystal detection facilitated data collection. Such strategy permitted collection of multiple crystals of radiation-free-damage data, resulting in the highest resolution of wild type (WT) Pol II crystals ever observed.
Keywords	DNA-directed RNA polymerase ; crystallization ; data collection ; monitoring ; nanocrystals ; transcription (genetics) ; transmission electron microscopy
Language	English
Dates of publication	2019-0415
Size	p. 23-28.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	1066584-5
ISSN	1095-9130 ; 1046-2023
ISSN (online)	1095-9130
ISSN	1046-2023
DOI	10.1016/j.ymeth.2019.04.011
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Transcription with a laser: Radiation-damage-free diffraction of RNA Polymerase II crystals.

Lin, Guowu / Weiss, Simon C / Vergara, Sandra / Camacho, Carlos / Calero, Guillermo

Methods (San Diego, Calif.)

2019 Volume 159-160, Page(s) 23–28

Abstract	Well-diffracting crystals are essential to obtain relevant structural data that will lead to understanding of RNA Polymerase II (Pol II) transcriptional processes at a molecular level. Here we present a strategy to study Pol II crystals using negative stain transmission electron microscopy (TEM) and a methodology to optimize radiation damage free data collection using free electron laser (FEL) at the Linac Coherent Light Source (LCLS). The use of negative stain TEM allowed visualization and optimization of crystal diffraction by monitoring the lattice quality of crystallization conditions. Nano crystals bearing perfect lattices were seeded and used to grow larger crystals for FEL data collection. Moreover, the use of in house designed crystal loops together with ultra-violet (UV) microscopy for crystal detection facilitated data collection. Such strategy permitted collection of multiple crystals of radiation-free-damage data, resulting in the highest resolution of wild type (WT) Pol II crystals ever observed.
MeSH term(s)	Crystallography/methods ; Humans ; Lasers ; Microscopy, Electron, Transmission/methods ; Models, Molecular ; Nanostructures ; Protein Conformation ; RNA Polymerase II/chemistry ; RNA Polymerase II/metabolism
Chemical Substances	RNA Polymerase II (EC 2.7.7.-)
Language	English
Publishing date	2019-04-25
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	1066584-5
ISSN	1095-9130 ; 1046-2023
ISSN (online)	1095-9130
ISSN	1046-2023
DOI	10.1016/j.ymeth.2019.04.011
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Article ; Online: The crystal structure of dGTPase reveals the molecular basis of dGTP selectivity.

Barnes, Christopher O / Wu, Ying / Song, Jinhu / Lin, Guowu / Baxter, Elizabeth L / Brewster, Aaron S / Nagarajan, V / Holmes, Andrew / Soltis, S Michael / Sauter, Nicholas K / Ahn, Jinwoo / Cohen, Aina E / Calero, Guillermo

Proceedings of the National Academy of Sciences of the United States of America

2019 Volume 116, Issue 19, Page(s) 9333–9339

Abstract: Deoxynucleotide triphosphohydrolases (dNTPases) play a critical role in cellular survival and DNA replication through the proper maintenance of cellular dNTP pools. While the vast majority of these enzymes display broad activity toward canonical dNTPs, ... ...

Abstract	Deoxynucleotide triphosphohydrolases (dNTPases) play a critical role in cellular survival and DNA replication through the proper maintenance of cellular dNTP pools. While the vast majority of these enzymes display broad activity toward canonical dNTPs, such as the dNTPase SAMHD1 that blocks reverse transcription of retroviruses in macrophages by maintaining dNTP pools at low levels,
MeSH term(s)	Allosteric Site ; Binding Sites ; Catalytic Domain ; Crystallography, X-Ray ; Deoxyguanine Nucleotides/chemistry ; Deoxyguanine Nucleotides/metabolism ; Escherichia coli/chemistry ; Escherichia coli/enzymology ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Escherichia coli Proteins/chemistry ; Escherichia coli Proteins/genetics ; Escherichia coli Proteins/metabolism ; GTP Phosphohydrolases/chemistry ; GTP Phosphohydrolases/genetics ; GTP Phosphohydrolases/metabolism ; Models, Molecular ; SAM Domain and HD Domain-Containing Protein 1/chemistry ; SAM Domain and HD Domain-Containing Protein 1/genetics ; SAM Domain and HD Domain-Containing Protein 1/metabolism ; Substrate Specificity
Chemical Substances	Deoxyguanine Nucleotides ; Escherichia coli Proteins ; deoxyguanosine triphosphate (8C2O37Y44Q) ; SAM Domain and HD Domain-Containing Protein 1 (EC 3.1.5.-) ; GTP Phosphohydrolases (EC 3.6.1.-)
Language	English
Publishing date	2019-04-24
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	209104-5
ISSN	1091-6490 ; 0027-8424
ISSN (online)	1091-6490
ISSN	0027-8424
DOI	10.1073/pnas.1814999116
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Zs.A 1148: Show issues

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Article ; Online: (N-{[4-(1,3-benzothiazol-2-yl)anilino]carbonylmethyl-kappaO}iminodiacetato-kappa(3)O,N,O')(1,10-phenanthroline-kappa(2)N,N')cobalt(II) pentahydrate.

Lin, Guo-Wu / Wang, Yue / Lu, Tao

Acta crystallographica. Section C, Crystal structure communications

2008 Volume 64, Issue Pt 4, Page(s) m179–81

Abstract: The title compound, [Co(C(19)H(15)N(3)O(5)S)(C(12)H(8)N(2))] x 5 H(2)O, has a moderately distorted octahedral coordination environment composed of two N atoms of a 1,10-phenanthroline ligand and one N and three O atoms of an N-{[4-(1,3-benzothiazol-2-yl) ... ...

Abstract	The title compound, [Co(C(19)H(15)N(3)O(5)S)(C(12)H(8)N(2))] x 5 H(2)O, has a moderately distorted octahedral coordination environment composed of two N atoms of a 1,10-phenanthroline ligand and one N and three O atoms of an N-{[4-(1,3-benzothiazol-2-yl)anilino]carbonylmethyl}iminodiacetate (ZL-5(2-)) ligand. The ring systems of the phenanthroline and ZL-5(2-) ligands are coplanar and the complexes pack in layers parallel to the ab plane with the rings of adjacent complexes facing one another. The layers stack along the c axis and are linked by hydrogen bonds involving the five water solvent molecules in the asymmetric unit and O atoms of the acetate groups of the ZL-5(2-) ligand. This is believed to be the first crystal structure of a complex of a 2-(4-aminophenyl)benzothiazole ligand.
MeSH term(s)	Cobalt/chemistry ; Crystallography ; Hydrogen Bonding ; Molecular Structure ; Phenanthrolines/chemistry
Chemical Substances	Phenanthrolines ; Cobalt (3G0H8C9362) ; 1,10-phenanthroline (W4X6ZO7939)
Language	English
Publishing date	2008-04
Publishing country	United States
Document type	Journal Article
ZDB-ID	2025703-X
ISSN	1600-5759 ; 2053-2296 ; 0108-2701
ISSN (online)	1600-5759 ; 2053-2296
ISSN	0108-2701
DOI	10.1107/S0108270108006306
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Article ; Online: Discovery of 4-((7H-Pyrrolo[2,3-d]pyrimidin-4-yl)amino)-N-(4-((4-methylpiperazin-1-yl)methyl)phenyl)-1H-pyrazole-3-carboxamide (FN-1501), an FLT3- and CDK-Kinase Inhibitor with Potentially High Efficiency against Acute Myelocytic Leukemia.

Wang, Yue / Zhi, Yanle / Jin, Qiaomei / Lu, Shuai / Lin, Guowu / Yuan, Haoliang / Yang, Taotao / Wang, Zhanwei / Yao, Chao / Ling, Jun / Guo, Hao / Li, Tonghui / Jin, Jianlin / Li, Baoquan / Zhang, Li / Chen, Yadong / Lu, Tao

Journal of medicinal chemistry

2018 Volume 61, Issue 4, Page(s) 1499–1518

Abstract: A series of 1-H-pyrazole-3-carboxamide derivatives have been designed and synthesized that exhibit excellent FLT3 and CDK inhibition and antiproliferative activities. A structure-activity-relationship study illustrates that the incorporation of a ... ...

Abstract	A series of 1-H-pyrazole-3-carboxamide derivatives have been designed and synthesized that exhibit excellent FLT3 and CDK inhibition and antiproliferative activities. A structure-activity-relationship study illustrates that the incorporation of a pyrimidine-fused heterocycle at position 4 of the pyrazole is critical for FLT3 and CDK inhibition. Compound 50 (FN-1501), which possesses potent inhibitory activities against FLT3, CDK2, CDK4, and CDK6 with IC
MeSH term(s)	Amides/chemistry ; Amides/pharmacology ; Animals ; Antineoplastic Agents/chemical synthesis ; Antineoplastic Agents/pharmacology ; Cell Line, Tumor ; Cyclin-Dependent Kinases/antagonists & inhibitors ; Drug Discovery ; Heterografts ; Humans ; Leukemia, Myeloid, Acute/drug therapy ; Mice ; Neoplasms/drug therapy ; Protein Kinase Inhibitors/pharmacology ; Protein Kinase Inhibitors/therapeutic use ; Pyrazoles/chemistry ; Pyrazoles/pharmacology ; Structure-Activity Relationship ; fms-Like Tyrosine Kinase 3/antagonists & inhibitors
Chemical Substances	Amides ; Antineoplastic Agents ; Protein Kinase Inhibitors ; Pyrazoles ; fms-Like Tyrosine Kinase 3 (EC 2.7.10.1) ; Cyclin-Dependent Kinases (EC 2.7.11.22)
Language	English
Publishing date	2018-02-12
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	218133-2
ISSN	1520-4804 ; 0022-2623
ISSN (online)	1520-4804
ISSN	0022-2623
DOI	10.1021/acs.jmedchem.7b01261
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Zs.B 151: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular ab Jg. 2022: Lesesaal (EG)
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Article ; Online: A facile synthesis of 3-substituted 9H-pyrido[3,4-b]indol-1(2H)-one derivatives from 3-substituted beta-carbolines.

Lin, Guowu / Wang, Yue / Zhou, Qingfa / Tang, Weifang / Wang, Jian / Lu, Tao

Molecules (Basel, Switzerland)

2010 Volume 15, Issue 8, Page(s) 5680–5691

Abstract: A mild and efficient two-step synthesis of 3-substituted beta-carbolinone derivatives from 3-substituted beta-carboline in good yields is described. A possible reaction mechanism for the formation of the skeleton of beta-carbolin-1-one is proposed. The ... ...

Abstract	A mild and efficient two-step synthesis of 3-substituted beta-carbolinone derivatives from 3-substituted beta-carboline in good yields is described. A possible reaction mechanism for the formation of the skeleton of beta-carbolin-1-one is proposed. The structures of these compounds were established by IR, 1H-NMR, 13C-NMR, mass spectrometry and elemental analysis, as well as X-ray crystallographic analysis of 4-2 and 6-2.
MeSH term(s)	Carbolines/chemistry ; Chemistry, Organic/methods ; Crystallography, X-Ray ; Indoles/chemical synthesis ; Indoles/chemistry ; Molecular Conformation ; Time Factors
Chemical Substances	Carbolines ; Indoles
Language	English
Publishing date	2010-08-17
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1413402-0
ISSN	1420-3049 ; 1431-5165 ; 1420-3049
ISSN (online)	1420-3049
ISSN	1431-5165 ; 1420-3049
DOI	10.3390/molecules15085680
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Article ; Online: Novel 1H-pyrazole-3-carboxamide derivatives: synthesis, anticancer evaluation and identification of their DNA-binding interaction.

Lu, Yi / Ran, Ting / Lin, Guowu / Jin, Qiaomei / Jin, Jianling / Li, Hongmei / Guo, Hao / Lu, Tao / Wang, Yue

Chemical & pharmaceutical bulletin

2013 Volume 62, Issue 3, Page(s) 238–246

Abstract: Four novel 1H-pyrazole-3-carboxamide derivatives were synthesized, and their antiproliferative effect on cancer cells, kinase inhibition, and in particular, the DNA-binding interaction were investigated to interpret the antitumor mechanisms. A DNA minor ... ...

Abstract	Four novel 1H-pyrazole-3-carboxamide derivatives were synthesized, and their antiproliferative effect on cancer cells, kinase inhibition, and in particular, the DNA-binding interaction were investigated to interpret the antitumor mechanisms. A DNA minor groove binding model was developed, and the binding energy was predicted for the compounds. In consistence with the prediction, the binding ability was determined by the electronic absorption spectroscopy under physiological conditions for the compounds, and further verified by viscosity measurement. One compound 5-(3-cyclopropylureido)-N-[4-[(4-methylpiperazin-1-yl)methyl]phenyl]-1-H-pyrazole-3-carboxamide (pym-5) exerted the highest DNA-binding affinity (K(pym-5)=1.06×10(5) M(-1)). And it demonstrated more than 50% decrease of the emission intensity of the ethidium bromide-calf thymus DNA (EB-CT-DNA) complex in fluorescence spectra, suggesting that pym-5 could strongly affect the DNA conformation. Furthermore, pym-5 showed the cleavage activity upon the supercoiled plasmid pBR322 DNA in the pBR322 DNA cleavage assay. Our study suggests that DNA may serve as a potential target to these pyrazole derivatives.
MeSH term(s)	Animals ; Antineoplastic Agents/chemical synthesis ; Antineoplastic Agents/chemistry ; Antineoplastic Agents/pharmacology ; Cattle ; Cell Line, Tumor ; DNA/chemistry ; Drug Screening Assays, Antitumor ; Humans ; Pyrazoles/chemical synthesis ; Pyrazoles/chemistry ; Pyrazoles/pharmacology
Chemical Substances	Antineoplastic Agents ; Pyrazoles ; DNA (9007-49-2) ; calf thymus DNA (91080-16-9)
Language	English
Publishing date	2013-12-20
Publishing country	Japan
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	213307-6
ISSN	1347-5223 ; 0009-2363
ISSN (online)	1347-5223
ISSN	0009-2363
DOI	10.1248/cpb.c13-00676
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