A polyplex human saliva peptide histatin 5-grafted methoxy PEG-b-polycaprolactone polymersome for intelligent stimuli-oriented doxorubicin delivery

2021 ◽  
pp. 102958
Author(s):  
Hichem Moulahoum ◽  
Faezeh Ghorbanizamani ◽  
Ece Bayir ◽  
Suna Timur ◽  
Figen Zihnioglu
Keyword(s):  
Human Saliva ◽  
Histatin 5 ◽  
Doxorubicin Delivery

scholarly journals Zinc Binding by Histatin 5 Promotes Fungicidal Membrane Disruption in C. albicans and C. glabrata

2020 ◽  
Vol 6 (3) ◽  
pp. 124 ◽  
Author(s):  
Hannah L. Norris ◽  
Rohitashw Kumar ◽  
Chih Yean Ong ◽  
Ding Xu ◽  
Mira Edgerton
Keyword(s):  
High Performance ◽  
Opportunistic Pathogen ◽  
Human Saliva ◽  
Membrane Disruption ◽  
Initial Surface ◽  
Surface Binding ◽  
Histatin 5 ◽  
Peptide Assembly ◽  
Cell Association ◽  
Exciting Potential

Histatin 5 (Hst 5) is an antimicrobial peptide produced in human saliva with antifungal activity for opportunistic pathogen Candida albicans. Hst 5 binds to multiple cations including dimerization-inducing zinc (Zn2+), although the function of this capability is incompletely understood. Hst 5 is taken up by C. albicans and acts on intracellular targets under metal-free conditions; however, Zn2+ is abundant in saliva and may functionally affect Hst 5. We hypothesized that Zn2+ binding would induce membrane-disrupting pores through dimerization. Through the use of Hst 5 and two derivatives, P113 (AA 4-15 of Hst 5) and Hst 5ΔMB (AA 1-3 and 15-19 mutated to Glu), we determined that Zn2+ significantly increases killing activity of Hst 5 and P113 for both C. albicans and Candida glabrata. Cell association assays determined that Zn2+ did not impact initial surface binding by the peptides, but Zn2+ did decrease cell association due to active peptide uptake. ATP efflux assays with Zn2+ suggested rapid membrane permeabilization by Hst 5 and P113 and that Zn2+ affinity correlates to higher membrane disruption ability. High-performance liquid chromatography (HPLC) showed that the higher relative Zn2+ affinity of Hst 5 likely promotes dimerization. Together, these results suggest peptide assembly into fungicidal pore structures in the presence of Zn2+, representing a novel mechanism of action that has exciting potential to expand the list of Hst 5-susceptible pathogens.

Salivary Histatin 5 and its Similarities to the Other Antimicrobial Proteins in Human Saliva

2000 ◽  
Vol 14 (1) ◽  
pp. 16-21 ◽  
Author(s):  
M. Edgerton ◽  
S.E. Koshlukova
Keyword(s):  
Antimicrobial Activity ◽  
Oral Cavity ◽  
Human Saliva ◽  
Oral Bacteria ◽  
Defense System ◽  
Antimicrobial Proteins ◽  
Cationic Proteins ◽  
Histatin 5 ◽  
Host Defense System ◽  
Bacteria And Fungi

Non-immune salivary proteins-including lactoperoxidase, lysozyme, lactoferrin, and histatins-are key components of the innate host defense system in the oral cavity. Many antimicrobial proteins contain multiple functional domains, with the result that one protein may have more than one mechanism of antimicrobial activity. These domains may be separated by proteolytic cleavage, creating smaller proteins with functional antimicrobial activity in saliva as described for lysozyme, lactoferrin, and histatins. These small cationic proteins then exert cytotoxic activity to oral bacteria and fungi. Salivary histatin 5 initiates killing of C. albicans through binding to yeast membrane proteins and non-lytic release of cellular ATP. Extracellular ATP may then activate fungal ATP receptors to induce ultimate cell death. This mechanism for fungal cytotoxicity may be shared by other antimicrobial cationic proteins. Microbicidal domains of salivary and host innate proteins should be considered as potential therapeutic agents in the oral cavity.

scholarly journals Circadian Rhythms of Histatin 1, Histatin 3, Histatin 5, Statherin and Uric Acid in Whole Human Saliva Secretion

2002 ◽  
Vol 33 (2) ◽  
pp. 213-222 ◽  
Author(s):  
M. Castagnola ◽  
T. Cabras ◽  
G. Denotti ◽  
M.B. Fadda ◽  
G. Gambarini ◽  
...  
Keyword(s):  
Uric Acid ◽  
Circadian Rhythms ◽  
Human Saliva ◽  
Histatin 5 ◽  
Saliva Secretion

scholarly journals In Vitro Assessment of Antifungal Therapeutic Potential of Salivary Histatin-5, Two Variants of Histatin-5, and Salivary Mucin (MUC7) Domain 1

2000 ◽  
Vol 44 (6) ◽  
pp. 1485-1493 ◽  
Author(s):  
Hongsa Situ ◽  
Libuse A. Bobek
Keyword(s):  
Human Serum ◽  
Amphotericin B ◽  
Therapeutic Potential ◽  
Artificial Saliva ◽  
Human Saliva ◽  
Trypan Blue Exclusion ◽  
Histatin 5 ◽  
Fungal Strains ◽  
Salivary Mucin

ABSTRACT Human salivary histatin-5 (Hsn-5) is a 24-residue peptide that possesses potent antifungal activity in vitro. The MUC7gene encodes human salivary low-molecular-weight mucin (MG2). The candidacidal activity of MUC7 domain 1 (MUC7 D1, the N-terminal 51 amino acid residues of MUC7) in vitro has also been demonstrated. In this study, we have investigated the antifungal therapeutic potential of Hsn-5, its two variants, R12I/K17N and R12I/H21L, and MUC7 D1. First, these peptides were tested for activities against different clinically important fungi. We found them to possess broad-spectrum antifungal activities; specifically, most exhibited excellent in vitro activity against eight clinically important fungal strains tested, including Candida albicansand Candida glabrata and their azole-resistant counterparts and Cryptococcus neoformans and its amphotericin B-resistant counterpart. These findings also suggest that the mechanism of action of both Hsn-5 and MUC7 D1 for these fungi is different from that of amphotericin B or azole antifungal agents. Second, we examined the stability of these peptides in whole human saliva and human serum. In saliva, the Hsn-5 variants R12I/K17N and R12I/H21L and MUC7 D1 degraded at a lower rate than Hsn-5. In human serum, MUC7 D1 was also more stable than Hsn-5; both peptides were more stable in serum than in saliva. Third, we examined the cytotoxicity of these peptides using human erythrocytes and two human cell lines (KB and HSG). No (or very low) hemolytic activity was observed with any of the four peptides, even at the highest protein concentration tested (200 μM), while amphotericin B caused 100% hemolysis at only 12.5 μM. The toxic effects of Hsn-5 and MUC7 D1 toward KB and HSG cells were also much lower than that of amphotericin B as measured by trypan blue exclusion. Together, these findings indicate that the investigated peptides possess high antifungal therapeutic potential, in particular for the treatment of drug-resistant fungal strains associated with immunocompromised (particularly human immunodeficiency virus-infected) patients. The same peptides could also be used as components of artificial saliva for patients with salivary dysfunction.

scholarly journals Studies of the mechanism of human salivary histatin-5 candidacidal activity with histatin-5 variants and azole-sensitive and -resistant Candida species.

1997 ◽  
Vol 41 (10) ◽  
pp. 2224-2228 ◽  
Author(s):  
H Tsai ◽  
L A Bobek
Keyword(s):  
Recombinant Proteins ◽  
Antifungal Agents ◽  
Structural Similarity ◽  
Human Saliva ◽  
Therapeutic Agents ◽  
Histatin 5 ◽  
Modeling Analysis ◽  
Antifungal Peptides ◽  
Resistant Strains ◽  
Number Of Cells

Histatins are a group of small, cationic, antifungal peptides present in human saliva. A previous molecular modeling analysis suggested structural similarity between the Phe14-His15 and His18-His19 dipeptide sequences in histatin-5 (Hsn-5; a 24-amino-acid polypeptide) and the sequence of miconazole (one of the azole-based antifungal therapeutic agents), implying that the mechanisms of killing of Candida albicans by these two molecules may be similar. To further elaborate on this observation, we have produced two variants of Hsn-5 in which Phe14-His15 or His18-His19 dipeptide sequences were replaced by Ala-Ala (F14A/H15A and H18A/H19A) to eliminate the phenyl and imidazole rings of the side chains and assessed their candidacidal activities against C. albicans. In addition, we tested azole-resistant C. albicans and Candida glabrata strains for their susceptibilities to Hsn-5. Analysis of the purified recombinant proteins for their candidacidal activities indicated that both variants were significantly less effective (the molar concentrations required to kill half of the maximum number of cells [ED50s], approximately 67 and approximately 149 microM for F14A/H15A and H18A/H19A, respectively) than the unaltered Hsn-5 (ED50, approximately 8 microM) at killing C. albicans, suggesting that the two dipeptide sequences are important for the candidacidal activity of Hsn-5. Assessment of the candidacidal activity of Hsn-5 with the well-characterized azole-resistant strains of C. albicans and C. glabrata, however, suggested that the mode of action of histatins against Candida is distinct from that of azole-based antifungal agents because Hsn-5 kills both azole-sensitive and azole-resistant strains equally well.

scholarly journals Identification of histatins as tannin-binding proteins in human saliva

1995 ◽  
Vol 311 (1) ◽  
pp. 341-347 ◽  
Author(s):  
Q Yan ◽  
A Bennick
Keyword(s):  
Tannic Acid ◽  
Biological Activities ◽  
Biological Effects ◽  
Condensed Tannin ◽  
Normal Growth ◽  
Human Saliva ◽  
Proline Content ◽  
Salivary Proteins ◽  
Parotid Saliva ◽  
Histatin 5

Tannins have a number of detrimental biological effects and these include interference with normal growth and metabolism if they are present in the feed of various animals. Proline-rich proteins (PRPs) in saliva have been shown to provide protection against tannin, but little is known about the mechanism of protection and interaction of other salivary proteins with tannin. To identify tannin-binding human salivary proteins, parotid and submandibular/sublingual saliva samples were adsorbed with tannin. PRPs, and in particular a group of low-M(r) proteins, were readily precipitated by tannin. The low-M(r) proteins were purified from parotid saliva and demonstrated to be histatins, a family of well-characterized histidine-rich salivary proteins. The ability of synthetic histatin 5, as well as an acidic PRP (PRP-1) and gelatin to precipitate quebracho condensed tannin and tannic acid was determined. At pH 7.4 histatin 5 was the most effective precipitant of both condensed tannin and tannic acid and it also precipitated the largest amount of condensed tannin at pH 3.0, but the smallest amount of tannic acid at that pH. In contrast PRP-1 showed a greater ability to precipitate both condensed tannin and tannic acid at pH 3.0 than at pH 7.4. Under most circumstances histatin 5 was therefore more effective in precipitating tannins than proteins with high proline content which generally have been recognized as strong precipitants of tannin. Pre-incubation of tannic acid with alpha-amylase inhibited the enzyme, but addition of histatin 5 or the acidic PRP PIF-s protected amylase from inhibition by tannin. Similarly salivary proteins may protect other biological activities in the digestive tract from inhibition by dietary tannin.

scholarly journals Histatin 5 Metallopeptides and Their Potential against Candida albicans Pathogenicity and Drug Resistance

Biomolecules ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1209
Author(s):  
Gabriela Vieira Silva Zolin ◽  
Fauller Henrique da Fonseca ◽  
Carolina Reis Zambom ◽  
Saulo Santesso Garrido
Keyword(s):  
Candida Albicans ◽  
Systemic Infection ◽  
Resistance Mechanisms ◽  
Human Saliva ◽  
Antifungal Drugs ◽  
New Drugs ◽  
Correct Treatment ◽  
Histatin 5 ◽  
Effective Drugs ◽  
High Antifungal Activity

Usually caused by Candida albicans, buccal candidiasis begins with the morphological transition between yeast and hyphal cells. Over time and without the correct treatment, it can be disseminated through the bloodstream becoming a systemic infection with high mortality rates. C. albicans already shows resistance against antifungals commonly used in treatments. Therefore, the search for new drugs capable of overcoming antifungal resistance is essential. Histatin 5 (Hst5) is an antimicrobial peptide of the Histatin family, that can be found naturally in human saliva. This peptide presents high antifungal activity against C. albicans. However, Hst5 action can be decreased for interaction with enzymes and metal ions present in the oral cavity. The current work aims to bring a brief review of relevant aspects of the pathogenesis and resistance mechanisms already reported for C. albicans. In addition, are also reported here the main immune responses of the human body and the most common antifungal drugs. Finally, the most important aspects regarding Histatin 5 and the benefits of its interaction with metals are highlighted. The intention of this review is to show the promising use of Hst5 metallopeptides in the development of effective drugs.

scholarly journals Candidacidal effects of two antimicrobial peptides: histatin 5 causes small membrane defects, but LL-37 causes massive disruption of the cell membrane

2005 ◽  
Vol 388 (2) ◽  
pp. 689-695 ◽  
Author(s):  
Alice L. den HERTOG ◽  
Jan van MARLE ◽  
Henk A. van VEEN ◽  
Wim van't HOF ◽  
Jan G. M. BOLSCHER ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cell Membrane ◽  
Antimicrobial Peptides ◽  
Freeze Fracture ◽  
Human Saliva ◽  
Morphological Properties ◽  
Histatin 5 ◽  
Membrane Morphology ◽  
Morphological Defects ◽  
Freeze Fracture Electron Microscopy

The effects of antimicrobial peptides on artificial membranes have been well-documented; however, reports on the ultrastructural effects on the membranes of micro-organisms are relatively scarce. We compared the effects of histatin 5 and LL-37, two antimicrobial peptides present in human saliva, on the functional and morphological properties of the Candida albicans cell membrane. Fluorescence microscopy and immunogold transmission electron microscopy revealed that LL-37 remained associated with the cell wall and cell membrane, whereas histatin 5 transmigrated over the membrane and accumulated intracellularly. Freeze-fracture electron microscopy revealed that LL-37 severely affected the membrane morphology, resulting in the disintegration of the membrane bilayer into discrete vesicles, and an instantaneous efflux of small molecules such as ATP as well as larger molecules such as proteins with molecular masses up to 40 kDa. The effects of histatin 5 on the membrane morphology were less pronounced, but still resulted in the efflux of nucleotides. As the morphological defects induced by histatin 5 are much smaller than those induced by LL-37, but the efflux of nucleotides is similar at comparable candidacidal concentrations, we suggest that the loss of nucleotides plays an important role in the killing process.

Effects of histatin 5 and derived peptides on Candida albicans

2001 ◽  
Vol 356 (2) ◽  
pp. 361-368 ◽  
Author(s):  
Anita L. A. RUISSEN ◽  
Jasper GROENINK ◽  
Eva J. HELMERHORST ◽  
Els WALGREEN-WETERINGS ◽  
Wim van't HOF ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Mitochondrial Membrane ◽  
Cytoplasmic Membrane ◽  
Human Saliva ◽  
Mitochondrial Activity ◽  
Complete Protection ◽  
Destructive Effect ◽  
Histatin 5 ◽  
Killing Activity ◽  
Anti Fungal

Three anti-microbial peptides were compared with respect to their killing activity against Candida albicans and their ability to disturb its cellular and internal membranes. Histatin 5 is an anti-fungal peptide occurring naturally in human saliva, while dhvar4 and dhvar5 are variants of its active domain, with increased anti-microbial activity. dhvar4has increased amphipathicity compared with histatin 5, whereas dhvar5has amphipathicity comparable with that of histatin 5. All three peptides caused depolarization of the cytoplasmic and/or mitochondrial membrane, indicating membranolytic activity. For the variant peptides both depolarization and killing occurred at a faster rate. With FITC-labelled peptides, no association with the cytoplasmic membrane was observed, contradicting the formation of permanent transmembrane multimeric peptide pores. Instead, the peptides were internalized and act on internal membranes, as demonstrated with mitochondrion- and vacuole-specific markers. In comparison with histatin 5, the variant peptides showed a more destructive effect on mitochondria. Entry of the peptides and subsequent killing were dependent on the metabolic state of the cells. Blocking of the mitochondrial activity led to complete protection against histatin 5 activity, whereas that of dhvar4 was hardly affected and that of dhvar5 was affected only intermediately.

Use of OPLC for quantitation of HCHO, as the dimedone adduct, in human saliva in different dental pathologies

2002 ◽  
Vol 15 (1) ◽  
pp. 19-22 ◽  
Author(s):  
T. Katarzyna Różyło ◽  
Anna Żabińska ◽  
Ingrid Różyło-Kalinowska
Keyword(s):  
Human Saliva
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