EcDBS1R6: A novel cationic antimicrobial peptide derived from a signal peptide sequence

The coding sequence of the SUC2 locus was placed under the control of the constitutive ADH1 promoter and transcription terminator in a centromere-based yeast plasmid vector from which invertase is expressed in a Suc- strain of Saccharomyces cerevisiae. Mutants in the signal peptide sequence were produced by replacing this region of the gene with synthetic oligonucleotide cassettes containing mixtures of nucleotides at several positions. The mutants could be divided into three classes on the basis of the ability to secrete invertase. Class I mutants produced secreted invertase but in reduced amount. The class II mutant, 4-55B, also exhibited reduced a level of invertase, but a significant fraction of the enzyme was intracellular. Class III mutants were partially defective in translocation from the cytoplasm to the endoplasmic reticulum and produced enzymatically active, unglycosylated preinvertase in the cytoplasm. Class III mutant preinvertases were also defective in translocation across canine pancreas microsomes. These results suggested that the reduced level of invertase resulted from proteolytic degradation of inefficiently transported intermediates. Comparison of the sequences of the mutant signal peptides indicated that amino acids at the extreme amino terminus and adjacent to the cleavage site play a crucial role in the secretory process when combined with a mutation within the hydrophobic core.

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Cell-free synthesis of the hirudin variant 1 of the blood-sucking leech Hirudo medicinalis

Scientific Reports ◽

10.1038/s41598-020-76715-w ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Doreen A. Wüstenhagen ◽

Phil Lukas ◽

Christian Müller ◽

Simone A. Aubele ◽

Jan-Peter Hildebrandt ◽

...

Keyword(s):

Signal Peptide ◽

Peptide Sequence ◽

Bacterial Cells ◽

Recombinant Hirudin ◽

Signal Peptide Sequence ◽

Cell Lysates ◽

Blood Sucking ◽

A Cell ◽

Synthesis Approach ◽

Cell Free Protein Synthesis

AbstractSynthesis and purification of peptide drugs for medical applications is a challenging task. The leech-derived factor hirudin is in clinical use as an alternative to heparin in anticoagulatory therapies. So far, recombinant hirudin is mainly produced in bacterial or yeast expression systems. We describe the successful development and application of an alternative protocol for the synthesis of active hirudin based on a cell-free protein synthesis approach. Three different cell lysates were compared, and the effects of two different signal peptide sequences on the synthesis of mature hirudin were determined. The combination of K562 cell lysates and the endogenous wild-type signal peptide sequence was most effective. Cell-free synthesized hirudin showed a considerably higher anti-thrombin activity compared to recombinant hirudin produced in bacterial cells.

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Production of 12-hydroxy dodecanoic acid methyl ester using a signal peptide sequence-optimized transporter AlkL and a novel monooxygenase

Bioresource Technology ◽

10.1016/j.biortech.2019.121812 ◽

2019 ◽

Vol 291 ◽

pp. 121812 ◽

Cited By ~ 1

Author(s):

Hee-Wang Yoo ◽

Joonwon Kim ◽

Mahesh D. Patil ◽

Beom Gi Park ◽

Sung-yeon Joo ◽

...

Keyword(s):

Methyl Ester ◽

Signal Peptide ◽

Acid Methyl Ester ◽

Peptide Sequence ◽

Dodecanoic Acid ◽

Signal Peptide Sequence ◽

Acid Methyl

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A New IL6 Isoform in Chinese Soft-Shelled Turtle (Pelodiscus sinesis) Discovered: Its Regulation during Cold Stress and Infection

Biology ◽

10.3390/biology9050111 ◽

2020 ◽

Vol 9 (5) ◽

pp. 111

Author(s):

Zuobing Zhang ◽

Miao Tian ◽

Ruxin Song ◽

Xiao Xing ◽

Yong Fan ◽

...

Keyword(s):

Cold Stress ◽

Signal Peptide ◽

Genomic Sequence ◽

Adaptive Immune Response ◽

Environmental Water ◽

Peptide Sequence ◽

Signal Peptide Sequence ◽

Gene Characterization ◽

Peptide Gene ◽

Response To Temperature

The Chinese soft-shelled turtle (Pelodiscus sinesis) is a widely cultured commercial species in East and Southeast Asian countries. The turtles frequently suffer from acute cold stress during farming in China. Stress-induced factor such as Interleukin-6 (IL6) is a multifunctional molecule that plays important roles in innate and adaptive immune response. In the present study, we found that the turtle possessed two IL6 transcripts, where one IL6 transcript contained a signal peptide sequence (psIL6), while the other IL6 transcript (psIL6ns) possessed no such signal peptide gene. To test any differential expression of the two isoforms during temperature and microbial stress, turtles were adapted to optimal environmental water temperature (25 °C), stressed by acute cooling for 24 h, followed with the challenge of Aeromonas hydrophila (1.8 × 108 CFU) or Staphylococcus aureus (5.8 × 108 CFU). Gene characterization revealed that psIL6ns, a splicer without codons encoding a signal peptide and identical to the one predicted from genomic sequence, and psIL6, a splicer with codons encoding a signal peptide, were both present. Inducible expression was documented in primary spleen cells stimulated with ConA and poly I: C. The splenic and intestinal expression of psIL6ns and psIL6 was increased in response to temperature stress and bacterial infection.

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Characterization of PauB, a Novel Broad-Spectrum Plasminogen Activator from Streptococcus uberis

Journal of Bacteriology ◽

10.1128/jb.184.1.119-125.2002 ◽

2002 ◽

Vol 184 (1) ◽

pp. 119-125 ◽

Cited By ~ 16

Author(s):

Philip N. Ward ◽

James A. Leigh

Keyword(s):

Plasminogen Activator ◽

Signal Peptide ◽

Open Reading Frame ◽

Peptide Sequence ◽

Expression Cloning ◽

Signal Peptide Sequence ◽

Putative Gene ◽

Reading Frame ◽

Streptococcus Uberis ◽

Broad Specificity

ABSTRACT A bovine plasminogen activator of atypical molecular mass (∼45 kDa) from Streptococcus uberis strain SK880 had been identified previously (L. B. Johnsen, K. Poulsen, M. Kilian, and T. E. Petersen. Infect. Immun. 67:1072–1078, 1999). The strain was isolated from a clinical case of bovine mastitis. The isolate was found not to secrete PauA, a bovine plasminogen activator expressed by the majority of S. uberis strains. Analysis of the locus normally occupied by pauA revealed an absence of the pauA open reading frame. However, an alternative open reading frame was identified within the same locus. Sequence analysis of the putative gene suggested limited but significant homology to other plasminogen activators. A candidate signal peptide sequence and cleavage site were also identified. Expression cloning of DNA encoding the predicted mature protein (lacking signal peptide) confirmed that the open reading frame encoded a plasminogen activator of the expected size, which we have named PauB. Both native and recombinant forms of PauB displayed an unexpectedly broad specificity profile for bovine, ovine, equine, caprine, porcine, rabbit, and human plasminogen. Clinical and nonclinical field isolates from nine United Kingdom sites were screened for the pauB gene and none were identified as carrying it. Similarly, clinical isolates from 20 Danish herds were all found to encode PauA and not PauB. Therefore, PauB represents a novel but rare bacterial plasminogen activator which displays very broad specificity.

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Addition of a signal peptide sequence to the α 1D -adrenoceptor gene increases the density of receptors, as determined by [3 H]-prazosin binding in the membranes

British Journal of Pharmacology ◽

10.1038/sj.bjp.0706087 ◽

2005 ◽

Vol 144 (5) ◽

pp. 651-659 ◽

Cited By ~ 18

Author(s):

Ramona Petrovska ◽

Ivo Kapa ◽

Janis Klovins ◽

Helgi B Schiöth ◽

Staffan Uhlén

Keyword(s):

Signal Peptide ◽

Peptide Sequence ◽

Signal Peptide Sequence

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Signal Peptide of Potato PinII Enhances the Expression of Cry1Ac in Transgenic Tobacco

Acta Biochimica et Biophysica Sinica ◽

10.1093/abbs/36.8.553 ◽

2004 ◽

Vol 36 (8) ◽

pp. 553-558 ◽

Cited By ~ 7

Author(s):

Yun-Jun Liu ◽

Yuan Yuan ◽

Jun Zheng ◽

Ya-Zhong Tao ◽

Zhi-Gang Dong ◽

...

Keyword(s):

Transgenic Tobacco ◽

Signal Peptide ◽

Proteinase Inhibitor ◽

Peptide Sequence ◽

Fluorescent Detection ◽

Transgenic Tobacco Plants ◽

Tobacco Plants ◽

Signal Peptide Sequence ◽

Proteinase Inhibitor Ii ◽

Cry1ac Protein

Abstract The modified Cry1Ac was expressed in transgenic tobacco plants. To allow secretion of the Cry1Ac protein into the intercellular space, the signal peptide sequence of potato proteinase inhibitor II (pinII) was N-terminally fused to the Cry1Ac encoding region. Expression of Cry1Ac in transgenic tobacco plants was assayed with ELISA. The results showed that pinII signal peptide sequence enhanced the expression of Cry1Ac protein and led to the secretion of the Cry1Ac protein in transgenic tobacco plants. GFP gene was also fused to the signal peptide sequence and transformed to tobacco. The results of fluorescent detection showed that GFP had localized in the apoplast of transgenic plants.

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EcDBS1R6: A new broad-spectrum cationic antibacterial peptide derived from a signal peptide sequence

10.1101/869867 ◽

2019 ◽

Author(s):

William F. Porto ◽

Luz N. Irazazabal ◽

Vincent Humblot ◽

Evan F. Haney ◽

Suzana M. Ribeiro ◽

...

Keyword(s):

Signal Peptide ◽

Broad Spectrum ◽

Antimicrobial Agents ◽

Membrane Damage ◽

Membrane Integrity ◽

Peptide Sequence ◽

World Health ◽

Signal Peptides ◽

Microscopy Imaging ◽

Signal Peptide Sequence

ABSTRACTBacterial infections represent a major worldwide health problem, with an special highlight on Gram-negative bacteria, which were assigned by the World Health Organization (WHO) as the most critical priority for development of novel antimicrobial compounds. Antimicrobial peptides (AMPs) have been considered as potential alternative agents for treating these infections. Here we demonstrated the broad-spectrum activity of EcDBS1R6, a peptide derived from a signal peptide sequence of Escherichia coli that we previously turned into an AMP by making changes predicted through the Joker algorithm. Signal peptides are known to naturally interact with membranes; however, the modifications introduced by Joker made this peptide capable of killing bacteria. Membrane damage of the bacterial cells was observed by measuring membrane integrity using fluorescent probes and through scanning electron microscopy imaging. Structural analysis revealed that the C-terminus was unable to fold into an α-helix, indicating that the EcDBS1R6 antibacterial activity core was located at the N-terminus, corresponding to the signal peptide portion of the parent peptide. Therefore, the strategy of transforming signal peptides into AMPs seems to be promising and could be used for producing novel antimicrobial agents.

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