Systematic deletion and site-directed mutagenesis of FHVB2 establish the role of C-terminal amino acid residues in RNAi suppression

2010 ◽  
Vol 398 (2) ◽  
pp. 290-295 ◽  
Author(s):  
Gatikrushna Singh ◽  
Reshma Korde ◽  
Pawan Malhotra ◽  
Sunil Mukherjee ◽  
Raj K. Bhatnagar
Keyword(s):  
Amino Acid ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Amino Acid Residues ◽  
Terminal Amino Acid ◽  
Terminal Amino

scholarly journals Deletion of the Actin-Associated Tropomyosin Tpm3 Leads to Reduced Cell Complexity in Cultured Hippocampal Neurons—New Insights into the Role of the C-Terminal Region of Tpm3.1

Cells ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 715
Author(s):  
Tamara Tomanić ◽  
Claire Martin ◽  
Holly Stefen ◽  
Esmeralda Parić ◽  
Peter Gunning ◽  
...  
Keyword(s):  
Amino Acid ◽  
Hippocampal Neurons ◽  
Molecular Mechanisms ◽  
Growth Cones ◽  
Amino Acid Residues ◽  
Terminal Amino Acid ◽  
C Terminus ◽  
Primary Mouse ◽  
Terminal Amino

Tropomyosins (Tpms) have been described as master regulators of actin, with Tpm3 products shown to be involved in early developmental processes, and the Tpm3 isoform Tpm3.1 controlling changes in the size of neuronal growth cones and neurite growth. Here, we used primary mouse hippocampal neurons of C57/Bl6 wild type and Bl6Tpm3flox transgenic mice to carry out morphometric analyses in response to the absence of Tpm3 products, as well as to investigate the effect of C-terminal truncation on the ability of Tpm3.1 to modulate neuronal morphogenesis. We found that the knock-out of Tpm3 leads to decreased neurite length and complexity, and that the deletion of two amino acid residues at the C-terminus of Tpm3.1 leads to more detrimental changes in neurite morphology than the deletion of six amino acid residues. We also found that Tpm3.1 that lacks the 6 C-terminal amino acid residues does not associate with stress fibres, does not segregate to the tips of neurites, and does not impact the amount of the filamentous actin pool at the axonal growth cones, as opposed to Tpm3.1, which lacks the two C-terminal amino acid residues. Our study provides further insight into the role of both Tpm3 products and the C-terminus of Tpm3.1, and it forms the basis for future studies that aim to identify the molecular mechanisms underlying Tpm3.1 targeting to different subcellular compartments.

scholarly journals Involvement of the N-terminal part of cyclophilin B in the interaction with specific Jurkat T-cell binding sites

1996 ◽  
Vol 317 (2) ◽  
pp. 571-576 ◽  
Author(s):  
Christophe MARILLER ◽  
Bernard HAENDLER ◽  
Fabrice ALLAIN ◽  
Agnès DENYS ◽  
Geneviève SPIK
Keyword(s):  
Amino Acid ◽  
Human Peripheral Blood ◽  
Biological Fluids ◽  
Peripheral Blood Lymphocytes ◽  
Site Directed Mutagenesis ◽  
Amino Acid Residues ◽  
Terminal Amino Acid ◽  
Cyclophilin B ◽  
Jurkat T Cell ◽  
Terminal Amino

Cyclophilin B (CyPB) is secreted in biological fluids such as blood or milk and binds to a specific receptor present on the human lymphoblastic cell line Jurkat and on human peripheral blood lymphocytes. This study was intended to specify the areas of CyPB that are involved in the interaction with the receptor. A synthetic peptide corresponding to the first 24 N-terminal amino acid residues of CyPB was shown to specifically recognize the receptor. Moreover, modification of Arg18 of CyPB by p-hydroxyphenylglyoxal led to a dramatic loss of affinity for the receptor. However, when this residue was replaced by an alanine residue using site-directed mutagenesis, no modification of the binding properties was found, suggesting that Arg18 is not directly involved but is sufficiently close to the interaction site to interfere with the binding when modified. Competitive binding experiments using a chimaeric protein made up of the 24 N-terminal amino acid residues of CyPB fused to the cyclophilin A core sequence confirmed the involvement of this region of CyPB in receptor binding.

The role of C-terminal amino acid residues of a Δ6-fatty acid desaturase from blackcurrant

2013 ◽  
Vol 431 (4) ◽  
pp. 675-679 ◽  
Author(s):  
Li-Ying Song ◽  
Wan-Xiang Lu ◽  
Jun Hu ◽  
Wei-Bo Yin ◽  
Yu-Hong Chen ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Amino Acid ◽  
Fatty Acid Desaturase ◽  
Amino Acid Residues ◽  
Terminal Amino Acid ◽  
Terminal Amino

scholarly journals Production of Native-Type Streptoverticillium mobaraense Transglutaminase in Corynebacterium glutamicum

2003 ◽  
Vol 69 (5) ◽  
pp. 3011-3014 ◽  
Author(s):  
Masayo Date ◽  
Kei-ichi Yokoyama ◽  
Yukiko Umezawa ◽  
Hiroshi Matsui ◽  
Yoshimi Kikuchi
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Corynebacterium Glutamicum ◽  
Cleavage Site ◽  
Active Form ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Terminal Amino Acid ◽  
Terminal Amino ◽  
Terminal Amino Acid Sequence

ABSTRACT We previously observed secretion of active-form transglutaminase in Corynebacterium glutamicum by coexpressing the subtilisin-like protease SAM-P45 from Streptomyces albogriseolus to process the prodomain. However, the N-terminal amino acid sequence of the transglutaminase differed from that of the native Streptoverticillium mobaraense enzyme. In the present work we have used site-directed mutagenesis to generate an optimal SAM-P45 cleavage site in the C-terminal region of the prodomain. As a result, native-type transglutaminase was secreted.

scholarly journals Identification of amino acid residues responsible for differences in substrate specificity and inhibitor sensitivity between two human liver dihydrodiol dehydrogenase isoenzymes by site-directed mutagenesis

1997 ◽  
Vol 323 (1) ◽  
pp. 61-64 ◽  
Author(s):  
Kazuya MATSUURA ◽  
Yoshihiro DEYASHIKI ◽  
Kumiko SATO ◽  
Naoko ISHIDA ◽  
Gunpei MIWA ◽  
...  
Keyword(s):  
Amino Acid ◽  
Human Liver ◽  
Hydroxysteroid Dehydrogenase ◽  
The Other ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Amino Acid Residues ◽  
Dihydrodiol Dehydrogenase ◽  
Inactive Protein

Human liver dihydrodiol dehydrogenase isoenzymes (DD1 and DD2), in which only seven amino acid residues are substituted, differ remarkably in specificity for steroidal substrates and inhibitor sensitivity: DD1 shows 20α-hydroxysteroid dehydrogenase activity and sensitivity to 1,10-phenanthroline, whereas DD2 oxidizes 3α-hydroxysteroids and is highly inhibited by bile acids. In the present study we performed site-directed mutagenesis of the seven residues (Thr-38, Arg-47, Leu-54, Cys-87, Val-151, Arg-170 and Gln-172) of DD1 to the corresponding residues (Val, His, Val, Ser, Met, His and Leu respectively) of DD2. Of the seven mutations, only the replacement of Leu-54 with Val produced an enzyme that had almost the same properties as DD2. No significant changes were observed in the other mutant enzymes. An additional site-directed mutagenesis of Tyr-55 of DD1 to Phe yielded an inactive protein, suggesting the catalytically important role of this residue. Thus a residue at a position before the catalytic Tyr residue might play a key role in determining the orientation of the substrates and inhibitors.

scholarly journals The Role of the Carboxyl-terminal Amino Acid Residues inEscherichia coliDNA Topoisomerase III-mediated Catalysis

1996 ◽  
Vol 271 (15) ◽  
pp. 9039-9045 ◽  
Author(s):  
Hong Liang Zhang ◽  
Swati Malpure ◽  
Zhiyu Li ◽  
Hiroshi Hiasa ◽  
Russell J. DiGate
Keyword(s):  
Amino Acid ◽  
Amino Acid Residues ◽  
Terminal Amino Acid ◽  
Topoisomerase Iii ◽  
Terminal Amino ◽  
Carboxyl Terminal ◽  
Mediated Catalysis
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