Identification of Serpin Determinants of Specificity and Selectivity for Furin Inhibition through Studies of α1PDX (α1-Protease Inhibitor Portland)-Serpin B8 and Furin Active-site Loop Chimeras

Izaguirre, Gonzalo; Qi, Lixin; Lima, M. I.; Olson, Steven T.

doi:10.1074/jbc.m113.462804

Cited by 7 publications

(50 citation statements)

References 32 publications

(39 reference statements)

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“… also reported that cathepsin L degrades filaggrin but serpin B13 (Hurpin) as does serpin B3 inhibits it, which may be mitigating filaggrinolysis . Furin, a member of the subtilisin/kexin‐like mammalian proprotein convertases, is also involved the (pro)filaggrin processing and its inhibitor serpin B8 is increased in the cheek samples which may account for the lack of total (pro)filaggrin processing on the cheek compared with the PA site . The UCA and PCA generating enzymes, histidine ammonia‐lyase (3×) and gamma‐glutamyl cyclotransferase, respectively, were also increased together with arginase‐1 (3×), that contributes to urea formation .…”

Section: Discussionmentioning

confidence: 99%

The effect of photodamage on the female Caucasian facial stratum corneum corneome using mass spectrometry‐based proteomics

Voegeli

Monneuse²,

Schoop

et al. 2017

Intern J of Cosmetic Sci

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BACKGROUND:The effect of photodamage on facial stratum corneum (SC) is still poorly understood. OBJECTIVE: To describe the SC proteome from tape strippings of Caucasian SC from photoexposed cheek and photoprotected postauricular (PA) site, a global analysis of photodamage on the skin will be developed leading to a better understanding of keratinocyte signalling pathways and identification of new molecular targets for the treatment of photoaged skin. METHODS: Female Caucasian subjects had nine consecutive tape strippings taken from their cheeks and PA site. Proteins were extracted and the trypsin-digested peptides were analysed by nanochromatography coupled to a high-resolution mass spectrometer. Data-dependent acquisition allowed protein identification that was processed by Paragon algorithm of Protein Pilot software. RESULTS: Changes in the levels of epidermal differentiation proteins were apparent indicating poor epidermal differentiation and SC maturation (keratins, cornified envelope (CE) proteins) on photoexposed cheeks. Differences in protease-anti-protease balance were observed for corneodesmolysis (favouring desquamation) and filaggrinolysis (favouring reduced filaggrin processing). 12R-LOX, a CE maturation enzyme, was reduced in photodamaged skin but not transglutaminases. Changes in signal keratinocyte transduction pathway markers were demonstrated especially by reduced levels of downstream signalling markers such as calreticulin (unfolded protein response; UPR) and increased level of stratifin (target of rapamycin; mTOR). Evidence for impaired proteostasis was apparent by reduced levels of a key proteasomal subunit (subunit beta type-6). Finally, key antioxidant proteins were upregulated except catalase. CONCLUSION: Clear examples of poor keratinocyte differentiation and associated metabolic and signalling pathways together with reduced SC maturation were identified in photodamaged facial SC. Corneocyte immaturity was evident with changes in CE proteins. Particularly, the reduction in 12R-LOX is a novel finding in photodamaged skin and supports the lack of SC maturation. Moreover, filaggrinolysis was reduced, whereas corneodesmolysis was enhanced. From our results, we propose that there is a poor crosstalk between the keratinocyte endoplasmic reticulum UPR, proteasome network and autophagy machinery that possibly leads to impaired keratinocyte proteostasis. Superimposed on these aberrations is an apparently enhanced mTOR pathway that also contributes to reduced SC formation and maturation. Our results clearly indicate a corneocyte scaffold disorder in photodamaged cheek SC.R esum e CONTEXTE: Les effets des dommages dus a l'exposition a la lumiere sur le stratum corneum (SC) sont encore mal compris. OBJECTIFS: D ecrire le prot eome du SC issu de pr el evements par bandes adh esives provenant de la joue, expos ee a la lumi ere, et de la r egion post-auriculaire, prot eg ee de la lumi ere, de sujets caucasiens.A cette fin, une analyse globale des dommages cutan es dus a l'exposition a la lumi er...

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Section: Discussionmentioning

confidence: 99%

The effect of photodamage on the female Caucasian facial stratum corneum corneome using mass spectrometry‐based proteomics

Voegeli

Monneuse²,

Schoop

et al. 2017

Intern J of Cosmetic Sci

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“…A similar approach could be used for tick serpins, where the reactive center loop (RCL), which is responsible for serpin specificity, would be of tick origin, and the conserved serpin scaffold would be of human origin (Silverman et al, 2010; Whisstock et al, 2010). The concept of creating novel serpins by combining the RCL and scaffold from different serpins has been proven with the fusion of the furin inhibitor B8 and the mutant variant of α1-antitrypsin (Izaguirre et al, 2013, 2019). Furthermore, a novel extracellular inhibitor of human granzyme B was produced by combining mouse RCL and a human scaffold into a single chimera (Marcet-Palacios et al, 2015).…”

Section: Tick Proteins and Bioengineeringmentioning

confidence: 99%

The Use of Tick Salivary Proteins as Novel Therapeutics

et al. 2019

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The last three decades of research into tick salivary components have revealed several proteins with important pharmacological and immunological activities. Two primary interests have driven research into tick salivary secretions: the search for suitable pathogen transmission blocking or "anti-tick" vaccine candidates and the search for novel therapeutics derived from tick salivary components. Intensive basic research in the field of tick salivary gland transcriptomics and proteomics has identified several major protein families that play important roles in tick feeding and overcoming vertebrate antitick responses. Moreover, these families contain members with unrealized therapeutic potential. Here we review the major tick salivary protein families exploitable in medical applications such as immunomodulation, inhibition of hemostasis and inflammation. Moreover, we discuss the potential, opportunities, and challenges in searching for novel tick-derived drugs.

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“…143,144 However, transplanting the P6-P1 RCL residues from the natural furin inhibitor Serpin B8 into α 1 -AT improved the rate of furin inhibition twofold versus α 1 -PDX, and surprisingly 10-fold versus the original Serpin B8. 145 Selectivity toward other proprotein convertases could be tuned via additional mutations, by introducing P 0 RCL residues from Serpin B8 or Serpin B8 exosite residues. 146 α 1 -AT, therefore, appears to be an attractive and tunable scaffold for engineering inhibitors of a variety of proprotein convertases.…”

Section: Inhibition Of Other Proteinasesmentioning

confidence: 99%

Engineering the serpin α₁‐antitrypsin: A diversity of goals and techniques

Scott

Sheffield

2019

Protein Science

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α1‐Antitrypsin (α1‐AT) serves as an archetypal example for the serine proteinase inhibitor (serpin) protein family and has been used as a scaffold for protein engineering for >35 years. Techniques used to engineer α1‐AT include targeted mutagenesis, protein fusions, phage display, glycoengineering, and consensus protein design. The goals of engineering have also been diverse, ranging from understanding serpin structure–function relationships, to the design of more potent or more specific proteinase inhibitors with potential therapeutic relevance. Here we summarize the history of these protein engineering efforts, describing the techniques applied to engineer α1‐AT, specific mutants of interest, and providing an appended catalog of the >200 α1‐AT mutants published to date.

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Identification of Serpin Determinants of Specificity and Selectivity for Furin Inhibition through Studies of α1PDX (α1-Protease Inhibitor Portland)-Serpin B8 and Furin Active-site Loop Chimeras

Cited by 7 publications

References 32 publications

The effect of photodamage on the female Caucasian facial stratum corneum corneome using mass spectrometry‐based proteomics

The effect of photodamage on the female Caucasian facial stratum corneum corneome using mass spectrometry‐based proteomics

The Use of Tick Salivary Proteins as Novel Therapeutics

Engineering the serpin α₁‐antitrypsin: A diversity of goals and techniques

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Identification of Serpin Determinants of Specificity and Selectivity for Furin Inhibition through Studies of α1PDX (α1-Protease Inhibitor Portland)-Serpin B8 and Furin Active-site Loop Chimeras

Cited by 7 publications

References 32 publications

The effect of photodamage on the female Caucasian facial stratum corneum corneome using mass spectrometry‐based proteomics

The effect of photodamage on the female Caucasian facial stratum corneum corneome using mass spectrometry‐based proteomics

The Use of Tick Salivary Proteins as Novel Therapeutics

Engineering the serpin α1‐antitrypsin: A diversity of goals and techniques

Contact Info

Product

Resources

About

Engineering the serpin α₁‐antitrypsin: A diversity of goals and techniques