Amino acid sequence determinants in self-assembly of insulin chiral amyloid superstructures: Role of C-terminus of B-chain in association of fibrils

Terminal deoxynucleotidyltransferase exists in multiple Mr forms, all apparently generated from a single polypeptide of 62kDa. On isolation and purification, the smallest catalytically active protein of this enzyme consists of two subunits, alpha (12kDa) and beta (30kDa). Recently a complementary-DNA nucleotide sequence has been reported for a portion of the enzyme from human lymphoblast. We have pinpointed the locations of the alpha- and beta-subunits within the elucidated nucleotide sequence. From these data, the portions of the nucleotide sequence coding for the catalytically important area of the transferase can be estimated. Here the amino acid sequence of a number of tryptic peptides from calf alpha- and beta-subunits is presented. Because of the striking homology between the amino acid sequence of the calf enzyme and that predicted for human lymphoblast enzyme, it is possible for us to conclude that the alpha-subunit was generated from the C-terminus of the precursor protein and the beta-subunit was non-overlapping and proximal.

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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 ◽

10.3390/cells10030715 ◽

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.

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CRISPR/Cas9-mediated mutation revealed cytoplasmic tail is dispensable for IZUMO1 function and male fertility

Reproduction ◽

10.1530/rep-16-0150 ◽

2016 ◽

Vol 152 (6) ◽

pp. 665-672 ◽

Cited By ~ 11

Author(s):

Samantha A M Young ◽

Haruhiko Miyata ◽

Yuhkoh Satouh ◽

Masanaga Muto ◽

Martin R Larsen ◽

...

Keyword(s):

Amino Acid ◽

Point Mutation ◽

Male Fertility ◽

Stop Codon ◽

Premature Stop Codon ◽

Cytoplasmic Tail ◽

Binding Partner ◽

C Terminus ◽

Manipulation System

IZUMO1 is a protein found in the head of spermatozoa that has been identified as essential for sperm–egg fusion. Its binding partner in the egg has been discovered (JUNO); however, the roles of several domains within IZUMO1 remain unexplored. One such domain is the C-terminus, which undergoes major phosphorylation changes in the cytoplasmic portion of the protein during rat epididymal transit. However, the cytoplasmic tail of IZUMO1 in many species is highly variable, ranging from 55 to one amino acid. Therefore, to understand the role of the cytoplasmic tail of IZUMO1 in mouse, we utilised the gene manipulation system of CRISPR/Cas9 to generate a point mutation resulting in a premature stop codon, producing mice with truncated IZUMO1. Mice without the cytoplasmic tail of IZUMO1 showed normal fertility but decreased the amount of protein, indicating that whilst this region is important for the expression level of IZUMO1, it is dispensable for fertilisation in the mouse.

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The amino acid sequence of yeast phosphoglycerate mutase

Proceedings of the Royal Society of London Series B Biological Sciences ◽

10.1098/rspb.1982.0026 ◽

1982 ◽

Vol 215 (1198) ◽

pp. 19-44 ◽

Cited By ~ 21

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Proteolytic Enzymes ◽

Phosphoglycerate Mutase ◽

Crystallographic Structure ◽

X Ray ◽

C Terminus ◽

Detailed Interpretation ◽

High Degree

The complete amino acid sequence of yeast phosphoglycerate mutase comprising 241 residues has been determined. The sequence was deduced from the two cyanogen bromide fragments, and from the peptides derived from these fragments after digestion by a number of proteolytic enzymes. Determination of this sequence now allows a detailed interpretation of the existing high-resolution X-ray crystallographic structure. A comparison of the sequence reported here with the sequences of peptides from phosphoglycerate mutases from other species, and with the sequence of erythrocyte diphosphoglycerate mutase, indicates that these enzymes have a high degree of structural homology. Autolysis of phosphoglycerate mutase by yeast extracts leads to the complete loss of mutase activity, and the formation of electrophoretically distinguishable forms (R. Sasaki, E. Sugimoto & H. Chiba, Archs Biochem. Biophys. 115, 53-61 (1966)). It is apparent from the amino acid sequence that these changes are due to the loss of an 8─12 residue peptide from the C-terminus.

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Assembly of amino acid containing naphthalene diimide-based molecules: the role of intervening amide groups in self-assembly, gelation, optical and semiconducting properties

Soft Matter ◽

10.1039/c8sm02460j ◽

2019 ◽

Vol 15 (14) ◽

pp. 3018-3026 ◽

Cited By ~ 10

Author(s):

Nibedita Nandi ◽

Kousik Gayen ◽

Arindam Banerjee

Keyword(s):

Amino Acid ◽

Self Assembly ◽

Naphthalene Diimide ◽

Semiconducting Properties

Two isomeric amino-acid containing naphthalene diimide based molecules differ in their respective assembly and the intervening amide groups plays an important role in gelation, optical and semiconducting nature of these molecules.

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Self-assembly and intracellular delivery of DNA by a truncated fragment derived from the Trojan peptide Penetratin

Soft Matter ◽

10.1039/d0sm00347f ◽

2020 ◽

Vol 16 (20) ◽

pp. 4746-4755 ◽

Cited By ~ 2

Author(s):

Lucas R. Mello ◽

Ian W. Hamley ◽

Valeria Castelletto ◽

Bianca B. M. Garcia ◽

Thiago C. Lourenço ◽

...

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Self Assembly ◽

Intracellular Delivery

A heptamer containing the longest noncationic amino acid sequence of Penetratin is shown to form peptiplexes with DNA. The structure is characterized from molecular to nanoscopic scale and peptiplexes are shown to assist intracell delivery of DNA.

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Purification, characterization, gene sequence, and significance of a bacterioferritin from Mycobacterium leprae.

Journal of Experimental Medicine ◽

10.1084/jem.180.1.319 ◽

1994 ◽

Vol 180 (1) ◽

pp. 319-327 ◽

Cited By ~ 57

Author(s):

M C Pessolani ◽

D R Smith ◽

B Rivoire ◽

J McCormick ◽

S A Hefta ◽

...

Keyword(s):

Amino Acid ◽

Membrane Protein ◽

Amino Acid Sequence ◽

Mycobacterium Leprae ◽

Native Molecular Mass ◽

Molecular Features ◽

Ferroxidase Center ◽

Considerable Homology

The study of tissue-derived Mycobacterium leprae provides insights to the immunopathology of leprosy and helps identify broad molecular features necessary for mycobacterial parasitism. A major membrane protein (MMP-II) of in vivo-derived M. leprae previously recognized (Hunter, S.W., B. Rivoire, V. Mehra, B.R. Bloom, and P.J. Brennan. 1990. J. Biol. Chem. 265:14065) was purified from extracts of the organism and partial amino acid sequence obtained. This information allowed recognition, within one of the cosmids that encompass the entire M. leprae genome, of a complete gene, bfr, encoding a protein of subunit size 18.2 kD. The amino acid sequence deduced from the major membrane protein II (MMP-II) gene revealed considerable homology to several bacterioferritins. Analysis of the native protein demonstrated the iron content, absorption spectrum, and large native molecular mass (380 kD) of several known bacterioferritins. The ferroxidase-center residues typical of ferritins were conserved in the M. leprae product. Oligonucleotides derived from the amino acid sequence of M. leprae bacterioferritin enabled amplification of much of the MMP-II gene and the detection of homologous sequences in Mycobacterium paratuberculosis, Mycobacterium avium, Mycobacterium tuberculosis, Mycobacterium intracellulare, and Mycobacterium scrofulaceum. The role of this iron-rich protein in the virulence of M. leprae is discussed.

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Mimicking the Function of Eggshell Matrix Proteins: The Role of Multiplets of Charged Amino Acid Residues and Self-Assembly of Peptides in Biomineralization

Angewandte Chemie International Edition ◽

10.1002/anie.200500261 ◽

2005 ◽

Vol 44 (34) ◽

pp. 5476-5479 ◽

Cited By ~ 27

Author(s):

Parayil Kumaran Ajikumar ◽

Subramanian Vivekanandan ◽

Rajamani Lakshminarayanan ◽

Seetharama D. S. Jois ◽

R. Manjunatha Kini ◽

...

Keyword(s):

Amino Acid ◽

Self Assembly ◽

Matrix Proteins ◽

Amino Acid Residues ◽

Eggshell Matrix

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The amino acid sequence of cytochrome c′ from Alcaligenes sp. N.C.I.B. 11015

Biochemical Journal ◽

10.1042/bj1350751 ◽

1973 ◽

Vol 135 (4) ◽

pp. 751-758 ◽

Cited By ~ 35

Author(s):

R. P. Ambler

Keyword(s):

Amino Acid ◽

Cytochrome C ◽

Amino Acid Sequence ◽

Attachment Site ◽

Photosynthetic Bacterium ◽

Chromatium Vinosum ◽

British Library ◽

Pseudomonas Denitrificans ◽

Single Polypeptide Chain ◽

C Terminus

The amino acid sequence of the cytochrome c′ from Alcaligenes sp. N.C.I.B. 11015 (Iwasaki's ‘Pseudomonas denitrificans’) has been determined. This organism is the only non-photosynthetic bacterium in which the protein has been found. The protein consists of a single polypeptide chain of 127 residues, with a single haem covalently attached to two cysteines. Unlike normal cytochromes c, the haem attachment site is very close to the C-terminus. The amino acid sequence around the haem attachment site is very similar to that of Chromatium vinosum D cytochrome c′. Detailed evidence for the amino acid sequence of the protein has been deposited as Supplementary Publication SUP 50022 at the British Library (Lending Division), (formerly the National Lending Library for Science and Technology), Boston Spa, Yorks. LS23 7BQ, U.K., from whom copies may be obtained on the terms given in Biochem. J. (1973) 131, 5.

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