The amino acid sequence of S-antigen: N-terminus and uveitogenic peptides

The preS1 surface glycoprotein of hepatitis B virus is targeted to the endoplasmic reticulum (ER) and is retained in this organelle when expressed in the absence of other viral gene products. The protein is also acylated at its N terminus with myristic acid. Sequences responsible for its ER retention have been identified through examination of mutants bearing lesions in the preS1 coding region. These studies reveal that such sequences map to the N terminus of the molecule, between residues 6 and 19. Molecules in which this region was present remained in the ER; those in which it had been deleted were secreted from the cell. Although all deletions which allowed efficient secretion also impaired acylation of the polypeptide, myristylation alone was not sufficient for ER retention: point mutations which eliminated myristylation did not lead to secretion. These data indicate that an essential element for ER retention resides in a 14-amino-acid sequence that is unrelated to previously described ER retention signals.

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Non-AUG Translation Initiation in a Plant RNA Virus: a Forty-Amino-Acid Extension Is Added to the N Terminus of the Soil-Borne Wheat Mosaic Virus Capsid Protein

Journal of Virology ◽

10.1128/jvi.72.2.1677-1682.1998 ◽

1998 ◽

Vol 72 (2) ◽

pp. 1677-1682 ◽

Cited By ~ 33

Author(s):

Yukio Shirako

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Mosaic Virus ◽

Capsid Protein ◽

Translation Initiation ◽

Deletion Mutant ◽

Rna Virus ◽

The United States ◽

Initiation Codon ◽

N Terminus

ABSTRACT RNA 2 of soil-borne wheat mosaic virus (SBWMV), the type species of the genus Furovirus, encodes a protein previously hypothesized to be initiated at an in-frame non-AUG codon upstream of the AUG initiation codon (nucleotide positions 334 to 336) for the 19-kDa capsid protein. Site-directed mutagenesis and in vitro transcription and translation analysis indicated that CUG (nucleotides 214 to 216) is the initiation codon for a protein with a calculated molecular mass of 25 kDa composed of a 40-amino-acid extension to the N terminus of the 19-kDa capsid protein. A stable deletion mutant, which was isolated after extensive passages of a wild-type SBWMV, contained a mixture of two deleted RNA 2’s, only one of which coded for the 25-kDa protein. The amino acid sequence of the N-terminal extension was moderately conserved and the CUG initiation codon was preserved among three SBWMV isolates from Japan and the United States. This amino acid sequence conservation, as well as the retention of expression of the 25-kDa protein in the stable deletion mutant, suggests that the 25-kDa protein is functional in the life cycle of SBWMV. This is the first report of a non-AUG translation initiation in a plant RNA virus genome.

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Amino acid sequence of the N-terminus and selected tryptic peptides of the active subunit of human plasma carboxypeptidase N: comparison with other carboxypeptidases

Biochemical and Biophysical Research Communications ◽

10.1016/0006-291x(88)90284-7 ◽

1988 ◽

Vol 154 (3) ◽

pp. 1323-1329 ◽

Cited By ~ 12

Author(s):

Randal A. Skidgel ◽

Carl D. Bennett ◽

James W. Schilling ◽

Fulong Tan ◽

Deepthi K. Weerasinghe ◽

...

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Human Plasma ◽

Tryptic Peptides ◽

N Terminus ◽

Carboxypeptidase N

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Selective Inactivation of Adrenomedullin over Calcitonin Gene-related Peptide Receptor Function by the Deletion of Amino Acids 14-20 of the Mouse Calcitonin-like Receptor

Journal of Biological Chemistry ◽

10.1074/jbc.m313058200 ◽

2004 ◽

Vol 279 (19) ◽

pp. 20387-20391 ◽

Cited By ~ 20

Author(s):

Daniela Koller ◽

Lars M. Ittner ◽

Roman Muff ◽

Knut Husmann ◽

Jan A. Fischer ◽

...

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Amino Acid Sequence ◽

Peptide Hormone ◽

Receptor Function ◽

Cgrp Receptor ◽

Related Peptide ◽

Calcitonin Gene ◽

N Terminus ◽

Camp Formation

The receptors for the neuropeptide calcitonin (CT) gene-related peptide (CGRP) and the multifunctional peptide hormone adrenomedullin (AM) are calcitonin-like receptor (CLR)/receptor-activity-modifying protein (RAMP) 1 and CLR/RAMP2 heterodimers, respectively. Here, the amino acid sequence TRNKIMT, corresponding to the residues 14-20 of the N terminus of the mouse (m) CLR, was found to be required for a functional mCLR/RAMP2 AM receptor. The deletion of amino acids 14-20 (Δ14-20) or their substitution by alanine (14-20A) did not affect the heterodimerization of the mCLR with mRAMP1 or mRAMP2, and the levels of expression at the surface of transiently transfected COS-7 cells were not altered. In mRAMP1/mCLR- or mRAMP1/mCLR-(Δ14-20)-expressing cells CGRP stimulated cAMP formation with EC50values of 0.12 ± 0.01 and 1.5 ± 0.4 nm, respectively. In mRAMP2/mCLR-expressing cells the EC50of AM was 0.8 ± 0.2 nm. However, in cells expressing mRAMP2/mCLR-(Δ14-20) up to 10-6mAM failed to stimulate cAMP production. In mRAMP2/mCLR-(14-20A) expressing cells the cAMP response to AM was minimally restored, and the EC50was >100 nm. In conclusion, the deletion of the amino acid sequence TRNKIMT of the extreme N terminus of the mCLR maintained CGRP receptor function of mRAMP1/receptor heterodimers, but AM no longer activated the mutant mCLR-(Δ14-20) in the presence of mRAMP2. The TRNKIMT sequence is required for normal mCLR/mRAMP2 association, and as a consequence, high affinity AM binding signaling the activation of adenylyl cyclase.

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The amino acid sequence of ferredoxin from the red alga Porphyra umbilicalis

Biochemical Journal ◽

10.1042/bj1730459 ◽

1978 ◽

Vol 173 (2) ◽

pp. 459-466 ◽

Cited By ~ 22

Author(s):

I Takruri ◽

B G Haslett ◽

D Boulter ◽

P W Andrew ◽

L J Rogers

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Red Alga ◽

Phenyl Isothiocyanate ◽

Plant Type ◽

Considerable Similarity ◽

Porphyra Umbilicalis ◽

N Terminus ◽

Red Algal ◽

Cnbr Cleavage

The amino acid sequence of the ferrodoxin of Porphyra umbilicalis was determined by the dansyl-phenyl isothiocyanate method, on peptides obtained by tryptic, chymotryptic and thermolytic digestion of the protein or its CNBr-cleavage fragments. The molecule consists of 98 residues, has an unblocked N-terminus and shows considerable similarity with other plant-type ferredoxins. It is the first reported sequence of a red-algal ferredoxin.

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Determination of the immunogenic region in the LipL32 protein of Leptospira

Asia Pacific Journal of Molecular Biology and Biotechnology ◽

10.35118/apjmbb.2018.026.3.04 ◽

2018 ◽

pp. 27-33

Author(s):

Mohammad Iskandar Jumat ◽

Kenneth Francis Rodrigues ◽

Azlyna Laribe ◽

Rashidah Mohammad ◽

Timothy William ◽

...

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

High Specificity ◽

Full Length ◽

Immunogenic Region ◽

C Terminus ◽

Initial Symptoms ◽

N Terminus ◽

Low Sensitivity

Leptospirosis is a zoonotic disease caused by the pathogenic species of Leptospira. The initial symptoms include fever, myalgia, nausea, skin rash, chills, and headache, which can be misdiagnosed. LipL32 is the highly conserved and abundant outer membrane protein (OMP) of Leptospira, which is used as an antigen in serodiagnostic assays. We used three in silico methods to predict the immunodominant regions in the full-length LipL32 protein. We identified three regions, namely the N-terminus (NrLipL32, amino acid sequence 20th-120th), intermediate (amino acid sequence 120th-150th), and C-terminus (CrLipL32, amino acid sequence 160th-260th) regions. The full-length protein and two larger fragments were cloned into the pET22b plasmid and expressed in Escherichia coli BL21 (DE3). The purified proteins were used as antigens in an ELISA to detect Leptospira-specific antibodies. The CrLipL32 ELISA showed the highest sensitivity for IgM (73.3%) and IgG (65%), followed by the full-length rLipL32 ELISA (IgM 68% and IgG 60%). The full-length rLipL32 ELISA showed high specificity (IgM 85% and IgG 75%), followed by the NrLipL32 ELISA (IgM 75% and IgG 60%). The intermediate fragment showed very low sensitivity (IgM 17% and IgG 2%). The sensitivity of the rLipL32 ELISA could be enhanced by adding other OMPs of Leptospira.

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CCAAT/Enhancer-Binding Protein ε27 Antagonism of GATA-1 Transcriptional Activity Is Mediated by a Unique N-Terminal Repression Domain, Is Independent of Sumoylation, and Does Not Require but Is Enhanced by DNA-Binding.

Blood ◽

10.1182/blood.v104.11.1468.1468 ◽

2004 ◽

Vol 104 (11) ◽

pp. 1468-1468 ◽

Cited By ~ 1

Author(s):

Monika J. Stankiewicz ◽

Jian Du ◽

Steven J. Ackerman

Keyword(s):

Amino Acid ◽

Dna Binding ◽

Amino Acid Sequence ◽

Terminal Differentiation ◽

Developmentally Regulated ◽

Protein Protein Interaction ◽

Enhancer Binding Protein ◽

N Terminus ◽

Repressor Activity ◽

Secondary Granule

Abstract The CCAAT/enhancer binding protein epsilon (C/EBPε) is critical for the terminal differentiation and lineage-specific gene expression of granulocytes, and expression of C/EBPε32 and its shorter 27 kD and 14 kD isoforms is developmentally regulated during neutrophil granulocyte differentiation. We have defined a novel role for the unique 27 kD isoform (C/EBPε27) as a potent antagonist of GATA-1-mediated transactivation of the promoter of the gene encoding the eosinophil secondary granule protein, major basic protein (MBP) (Du et al, J. Biol. Chem.2002; 277:43481–43394). We also showed that these two transcription factors physically interact in eosinophil cell lines in vivo. In the present studies, we performed the first structure-function analyses of the C/EBPε27 isoform to map its potent repressor domains, with comparisons to the C/EBPε32 and C/EBPε14 isoforms, using transactivation assays of the MBP P2 promoter in the presence of GATA-1. Our results show that the repression of GATA-1 is mediated in part by the unique N-terminus of C/EBPε27 (not shared with other C/EBPε isoforms) in combination with part of a previously identified RDI domain (shared with full length C/EBPε32). We show further that this repressor activity is independent of DNA binding (via deletion of the basic region of C/EBPε27) as well as of sumoylation of the RDI “VKEEP” sumoylation consensus site present in both the C/EBPε32 and C/EBPε27 isoforms, and conserved in the C/EBPε proteins of many other species. Thus, our findings identify the unique N-terminus of the C/EBPε27 isoform, a distinct 68 amino acid sequence not shared with any other C/EBPε isoforms or other C/EBP family members, as the minimum repressor domain required for potent antagonism of GATA-1 activity. Of interest, fusion of this novel 68 amino acid sequence to the N-terminus of full length C/EBPε32 converted it into a partial repressor of GATA-1, but did not alter the transactivation potential of the C/EBPε32 isoform itself. The mechanism for maximal C/EBPε27 attenuation of GATA-1 activity requires a combination of both GATA-1-C/EBPε27 protein-protein interaction and C/EBPε27 binding to the proximal C/EBP consensus site immediately upstream in the target promoter. Neither C/EBPε32 nor C/EBPε14 inhibited C/EBPε27 antagonism of GATA-1, supporting a protein-protein interaction mechanism for its repressor activity that is enhanced by, but does not require, DNA binding to a proximal C/EBP site. Expression of the C/EBPε27 isoform likely serves to titrate and/or turn off expression of secondary granule protein genes such as MBP during eosinophil terminal differentiation, when these genes are ultimately silenced in the mature cell. These studies illustrate the unique regulatory (activating versus repressor) activities for the various C/EBPε isoforms, activities consistent with their developmentally regulated expression and lineage-specific activities during granulocyte (both neutrophil and eosinophil) differentiation.

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Partial amino acid sequence of rat pre-prolactin

Biochemical Journal ◽

10.1042/bj1610189 ◽

1977 ◽

Vol 161 (1) ◽

pp. 189-192 ◽

Cited By ~ 34

Author(s):

R A Maurer ◽

J Gorski ◽

D J McKean

Keyword(s):

Amino Acid ◽

Sequence Analysis ◽

Amino Acid Sequence ◽

Cysteine Residue ◽

Amino Acid Residues ◽

Partial Amino Acid Sequence ◽

Considerable Similarity ◽

Rat Pituitary ◽

N Terminus ◽

Methionine Residues

Rat pituitary mRNA was used to direct the cell-free synthesis of pre-prolactin labelled with [4,5-3H]leucine and either [35S] methioninc or [35S] cystine. Sequence analysis of the labelled protein indicates that pre-prolactin has 29 amino acid residues joined to the N-terminus of the prolactin sequence. Leucine residues were found at positions 13, 14, 15, 16, 21 and 22, methionine residues at positions 1, 17 and 18, and a cysteine residue at position 24 of the precursor sequence, and this partial sequence shows considerable similarity with other precursors that have been sequenced.

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Identification of N-terminal methionine in the precursor of immunoglobulin light chain. Initiation of translation of messenger ribonucleic acid in plants and animals

Biochemical Journal ◽

10.1042/bj1530543 ◽

1976 ◽

Vol 153 (3) ◽

pp. 543-550 ◽

Cited By ~ 23

Author(s):

I Schechter ◽

Y Burstein

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Light Chain ◽

Mrna Translation ◽

Mature Protein ◽

Free System ◽

Cell Free System ◽

Messenger Ribonucleic Acid ◽

Initiation Of Translation ◽

N Terminus

The proteins programmed in the wheat-germ cell-free system by the mRNA coding for the MOPC-321 mouse myeloma L (light) chain were labelled with [35S]methionine, [4,5-3H]leucine or [3-3H]serine, and were subjected to amino acid-sequence analyses. Over 95% of the total cell-free product was sequenced as one homogeneous protein, which corresponds to the precursor of the L-chain protein. In the precursor, 20 amino acid residues precede the N-terminus of the mature protein. This extra piece contains one methionine residue at the N-terminus, one serine residue at position 18, and six leucine residues, which are clustered in two triplets at positions 6, 7, 8 and 11, 12, 13. The identification of methionine at the N-terminus of the precursor is in agreement with the evidence showing that unblocked methionine is the initiator residue for protein synthesis in eukaryotes. The absence of methionine at position 20, which precedes the N-terminal residue of the mature protein, suggests that myeloma cells synthesize the precursor. However, within the cell the precursor should be rapidly processed to the mature L chain, since precursor molecules have not yet been found in the intact animal. The abundance (30%) of leucine residues indicates that the extra-piece moiety is quite hydrophobic. The extra piece of the MOPC-321 L-chain precursor synthesized with the aid of the Krebs II ascites cell-free system is of identical size and it has the same leucine sequence [Schechter et al. (1975) Science 188, 160-162]. This indicates that cell-free systems derived from the plant and animal kingdom initiate mRNA translation from the same point. It is shown that the amino acid sequence of minute amounts of a highly labelled protein (0.1 pmol) can be faithfully determined in the presence of a large excess (over 2000 000-fold) of unrelated non-radioactive proteins.

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