scholarly journals The N-terminal amino acid sequences of the heavy and light chains of human cathepsin L Relationship to a cDNA clone for a major cysteine proteinase from a mouse macrophage cell line

1986 ◽  
Vol 240 (2) ◽  
pp. 373-377 ◽  
Author(s):  
R W Mason ◽  
J E Walker ◽  
F D Northrop
Keyword(s):  
Amino Acids ◽  
Cell Line ◽  
Heavy Chain ◽  
Cdna Clone ◽  
Cysteine Proteinase ◽  
Cathepsin L ◽  
Amino Acid Sequences ◽  
Macrophage Cell ◽  
Terminal Amino ◽  
Mouse Macrophage Cell Line

Human liver cathepsin L consists of a heavy chain and a light chain with Mr values of 25,000 and 5000 respectively. The chains have been purified and their N-terminal amino acid sequences have been determined. The 40 amino acids determined from the heavy chain and 42 amino acids sequenced in the light chain are homologous with the N-terminal and C-terminal regions respectively of the superfamily of cysteine proteinases. Therefore it is likely that the two chains of cathepsin L are derived by proteolysis of a single polypeptide precursor. Of the amino acids sequenced, 81% are identical with the homologous portions of a protein sequence for a major cysteine proteinase predicted from a cDNA clone from a mouse macrophage cell line. This is the closest relative amongst the known sequences in the superfamily and strongly indicates that the protein encoded by this mRNA is cathepsin L. The mouse protein is also probably the major excreted protein of a transformed cell line [Gal & Gottesman (1986) Biochem. Biophys. Res. Commun. 139, 156-162]. The heavy chain is identical in only 71% of its residues with the sequence of ox cathepsin S, providing further evidence that this latter enzyme is probably not a species variant of cathepsin L. The relationship with a second unidentified cathepsin cDNA clone from a bovine library is much weaker (41% identity), and so this clone remains unidentified.

scholarly journals Roles of the C-Terminal Amino Acids of Non-Hexameric Helicases: Insights from Escherichia coli UvrD

2021 ◽  
Vol 22 (3) ◽  
pp. 1018
Author(s):  
Hiroaki Yokota
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Amino Acid ◽  
Single Molecule ◽  
Underlying Mechanism ◽  
Amino Acid Sequences ◽  
E Coli ◽  
X Ray Crystallography ◽  
Terminal Amino

Helicases are nucleic acid-unwinding enzymes that are involved in the maintenance of genome integrity. Several parts of the amino acid sequences of helicases are very similar, and these quite well-conserved amino acid sequences are termed “helicase motifs”. Previous studies by X-ray crystallography and single-molecule measurements have suggested a common underlying mechanism for their function. These studies indicate the role of the helicase motifs in unwinding nucleic acids. In contrast, the sequence and length of the C-terminal amino acids of helicases are highly variable. In this paper, I review past and recent studies that proposed helicase mechanisms and studies that investigated the roles of the C-terminal amino acids on helicase and dimerization activities, primarily on the non-hexermeric Escherichia coli (E. coli) UvrD helicase. Then, I center on my recent study of single-molecule direct visualization of a UvrD mutant lacking the C-terminal 40 amino acids (UvrDΔ40C) used in studies proposing the monomer helicase model. The study demonstrated that multiple UvrDΔ40C molecules jointly participated in DNA unwinding, presumably by forming an oligomer. Thus, the single-molecule observation addressed how the C-terminal amino acids affect the number of helicases bound to DNA, oligomerization, and unwinding activity, which can be applied to other helicases.

scholarly journals Proteome dataset of mouse macrophage cell line infected with tick-borne encephalitis virus

Data in Brief ◽  
2020 ◽  
Vol 28 ◽  
pp. 105029
Author(s):  
A.L. Rusanov ◽  
A.A. Stepanov ◽  
V.G. Zgoda ◽  
A.L. Kaysheva ◽  
M. Selinger ◽  
...  
Keyword(s):  
Cell Line ◽  
Encephalitis Virus ◽  
Mouse Macrophage ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Tick Borne Encephalitis ◽  
Tick Borne Encephalitis Virus ◽  
Mouse Macrophage Cell Line

scholarly journals Antileishmanial Activities of Several Classes of Aromatic Dications

2002 ◽  
Vol 46 (3) ◽  
pp. 797-807 ◽  
Author(s):  
James J. Brendle ◽  
Abram Outlaw ◽  
Arvind Kumar ◽  
David W. Boykin ◽  
Donald A. Patrick ◽  
...  
Keyword(s):  
Cell Line ◽  
Leishmania Donovani ◽  
Pneumocystis Carinii ◽  
Antileishmanial Activity ◽  
Microbial Pathogens ◽  
Mouse Macrophage ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Mouse Macrophage Cell Line

ABSTRACT Aromatic dicationic molecules possess impressive activity against a broad spectrum of microbial pathogens, including Pneumocystis carinii, Cryptosporidium parvum, and Candida albicans. In this work, 58 aromatic cations were examined for inhibitory activity against axenic amastigote-like Leishmania donovani parasites. In general, the most potent of the compounds were substituted diphenyl furan and thiophene dications. 2,5-Bis-(4-amidinophenyl)thiophene was the most active compound. This agent displayed a 50% inhibitory concentration (IC50) of 0.42 ± 0.08 μM against L. donovani and an in vitro antileishmanial potency 6.2-fold greater than that of the clinical antileishmanial dication pentamidine and was 155-fold more toxic to the parasites than to a mouse macrophage cell line. 2,4-Bis-(4-amidinopheny)furan was twice as active as pentamidine (IC50, 1.30 ± 0.21 μM), while 2,5-bis-(4-amidinopheny)furan and pentamidine were essentially equipotent in our in vitro antileishmanial assay. Carbazoles, dibenzofurans, dibenzothiophenes, and benzimidazoles containing amidine or substituted amidine groups were generally less active than the diphenyl furans and thiophenes. In all cases, aromatic dications possessing strong antileishmanial activity were severalfold more toxic to the parasites than to a cultured mouse macrophage cell line. These structure-activity relationships demonstrate the potent antileishmanial activity of several aromatic dications and provide valuable information for the future design and synthesis of more potent antiparasitic agents.

scholarly journals Effect of N- and C-Terminal Amino Acids on the Interfacial Binding Properties of Phospholipase D from Vibrio parahaemolyticus

2018 ◽  
Vol 19 (8) ◽  
pp. 2447
Author(s):  
Fanghua Wang ◽  
Ruixia Wei ◽  
Abdelkarim Abousalham ◽  
Wuchong Chen ◽  
Bo Yang ◽  
...  
Keyword(s):  
Amino Acids ◽  
Secondary Structure ◽  
Phospholipase D ◽  
Vibrio Parahaemolyticus ◽  
Terminal Segment ◽  
Amino Acid Sequences ◽  
Loop Structure ◽  
Binding Properties ◽  
Phospholipid Monolayers ◽  
Terminal Amino

The effects of N-terminal (1–34 amino acids) and C-terminal (434–487 amino acids) amino acid sequences on the interfacial binding properties of Phospholipase D from Vibrio parahaemolyticus (VpPLD) were characterized by using monomolecular film technology. Online tools allowed the prediction of the secondary structure of the target N- and C-terminal VpPLD sequences. Various truncated forms of VpPLD with different N- or C-terminal deletions were designed, based on their secondary structure, and their membrane binding properties were examined. The analysis of the maximum insertion pressure (MIP) and synergy factor “a” indicated that the loop structure (1–25 amino acids) in the N-terminal segment of VpPLD had a positive effect on the binding of VpPLD to phospholipid monolayers, especially to 1,2-dimyristoyl-sn-glycero-3-phosphoserine and 1,2-dimyristoyl-sn-glycero-3-phosphocholine. The deletion affecting the N-terminus loop structure caused a significant decrease of the MIP and synergy factor a of the protein for these phospholipid monolayers. Conversely, the deletion of the helix structure (26–34 amino acids) basically had no influence on the binding of VpPLD to phospholipid monolayers. The deletion of the C-terminal amino acids 434–487 did not significantly change the binding selectivity of VpPLD for the various phospholipid monolayer tested here. However, a significant increase of the MIP value for all the phospholipid monolayers strongly indicated that the three-strand segment (434–469 amino acids) had a great negative effect on the interfacial binding to these phospholipid monolayers. The deletion of this peptide caused a significantly greater insertion of the protein into the phospholipid monolayers examined. The present study provides detailed information on the effect of the N- and C-terminal segments of VpPLD on the interfacial binding properties of the enzyme and improves our understanding of the interactions between this enzyme and cell membranes.

scholarly journals Introduction of a human colony stimulating factor-1 gene into a mouse macrophage cell line induces CSF-1 independence but not tumorigenicity

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1218-1225 ◽  
Author(s):  
MF Roussel ◽  
CW Rettenmier ◽  
CJ Sherr
Keyword(s):  
Cell Line ◽  
Retroviral Vector ◽  
Gene Products ◽  
Mouse Macrophage ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Multiple Copies ◽  
Membrane Bound ◽  
Stimulating Factor ◽  
Mouse Macrophage Cell Line

Abstract The SV40-immortalized mouse macrophage cell line, BAC1.2F5, is strictly dependent on CSF-1 for its survival and proliferation in culture. Introduction of a retroviral vector containing a 1.6 kilobase (kb) pair human CSF-1 cDNA into these cells abrogated their growth factor dependence but did not render the cells tumorigenic in nude mice. The infected macrophages contained multiple copies of the vector provirus, expressed both membrane-bound and secreted forms of CSF-1, and exhibited constitutive down modulation of the murine CSF-1 receptor. Because insertion of the v-fms gene has previously been shown to abrogate factor dependence and induce tumorigenicity in BAC1.2F5 macrophages, the failure of these cells to express a fully transformed phenotype after persistent stimulation by endogenous CSF-1 suggests that the v-fms and c-fms gene products provide different signals for cell proliferation.

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