Human Calprotectin: Effect of Calcium and Zinc on its Secondary and Tertiary Structures, and Role of pH in its Thermal Stability

Yousefi, Reza; Imani, Mehdi; Ardestani, Sussan Kaboudanian; Saboury, Ali Akbar; Gheibi, Nematollah; Ranjbar, Bijian

doi:10.1111/j.1745-7270.2007.00343.x

Cited by 34 publications

(37 citation statements)

References 42 publications

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“…Purification and Use of Calprotectin from Human Neutrophils-Calprotectin was purified from the cytosol of human neutrophils obtained by nitrogen cavitation using a previously described method (14) with Coomassie Blue staining of reducing SDS-PAGE gels indicating that the purity of calprotectin was 93% with the major contaminant, S100A12, making up 6%. For mass spectrometry and protein carbonyl experiments, calprotectin (40 M) was incubated with HNE (0 -800 M) in PBS at 37°C for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Myeloperoxidase-dependent Lipid Peroxidation Promotes the Oxidative Modification of Cytosolic Proteins in Phagocytic Neutrophils

Wilkie-Grantham¹,

Magon²,

Harwood

et al. 2015

Journal of Biological Chemistry

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Background: Neutrophils generate oxidants to kill microbes ingested into phagosomes, but extraphagosomal protein oxidation may also occur. Results: Specific neutrophil cytosolic proteins were carbonylated following phagocytosis; carbonylation was impaired by myeloperoxidase inhibitors or radical scavengers that inhibit lipid peroxidation. Conclusion: Lipid peroxidation in the phagosomal membrane leads to oxidation of cytosolic proteins. Significance: Protein oxidation may influence the function and fate of post-phagocytic neutrophils.

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Section: Methodsmentioning

confidence: 99%

Myeloperoxidase-dependent Lipid Peroxidation Promotes the Oxidative Modification of Cytosolic Proteins in Phagocytic Neutrophils

Wilkie-Grantham¹,

Magon²,

Harwood

et al. 2015

Journal of Biological Chemistry

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“…CP inhibits bacterial growth and metal-dependent virulence factors by sequestering Mn and Zn ions. However, the relative contribution of Mn vs. Zn sequestration and the corresponding effects on specific cellular pathways are unknown (11,13,20). To address this issue, the ΔS1 and ΔS2 CP mutants were used to assess the individual contributions of Mn and Zn sequestration to inhibiting S. aureus growth.…”

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confidence: 99%

Molecular basis for manganese sequestration by calprotectin and roles in the innate immune response to invading bacterial pathogens

Damo

Kehl-Fie

Sugitani

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

340

594

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The S100A8/S100A9 heterodimer calprotectin (CP) functions in the host response to pathogens through a mechanism termed "nutritional immunity." CP binds Mn 2+ and Zn 2+ with high affinity and starves bacteria of these essential nutrients. Combining biophysical, structural, and microbiological analysis, we identified the molecular basis of Mn 2+ sequestration. The asymmetry of the CP heterodimer creates a single Mn 2+ -binding site from six histidine residues, which distinguishes CP from all other Mn 2+ -binding proteins. Analysis of CP mutants with altered metal-binding properties revealed that, despite both Mn 2+ and Zn 2+ being essential metals, maximal growth inhibition of multiple bacterial pathogens requires Mn 2+ sequestration. These data establish the importance of Mn 2+ sequestration in defense against infection, explain the broad-spectrum antimicrobial activity of CP relative to other S100 proteins, and clarify the impact of metal depletion on the innate immune response to infection.bacterial pathogenesis | Staphylococcus aureus | antibiotic resistance | protein crystal structure | isothermal titration calorimetry B acterial pathogens are a significant threat to global public health. This threat is compounded by the fact that these organisms are rapidly becoming resistant to all relevant antimicrobials. Of particular note is the recent emergence of antibiotic-resistant strains of Staphylococcus aureus as a leading cause of bacterial infection in the United States (1) and arguably the most important threat to the public health of the developed world. Consequently, the identification of therapeutics to treat bacterial pathogens is paramount to our continued ability to limit this infectious threat.One promising area of potential therapeutic development involves targeting bacterial access to essential transition metals. This strategy is based on the fact that all bacterial pathogens require these nutrient metals to colonize their hosts (2-5). In vertebrates, the bacterial need for nutrient transition metals is counteracted by the sequestration of these metals by the host. This limitation of essential nutrients, termed "nutritional immunity," is a potent defense against infection (6). Although a variety of metals is required for microbial growth, studies of nutritional immunity have been primarily restricted to the struggle for iron (Fe) between host and pathogen (7-9).An innate immune factor, calprotectin (CP), is abundant in neutrophils and plays a key role in nutritional immunity. CP can be found at sites of infection in excess of 1 mg/mL and is required for the control of a number of medically relevant bacteria and fungi including S. aureus, Candida albicans, and Aspergillus fumigates (10-14). The antimicrobial activity of CP is due to the chelation of the essential nutrients Zn 2+ (Zn) and Mn 2+ (Mn), which results in bacterial metal starvation and is reversed by the addition of these metals in excess (11, 13). Moreover, CP-deficient mice have increased microbial burdens following systemic challenge, und...

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“…In fact, examination of abscesses formed during S. aureus infection has revealed that these lesions are Mn and Zn depleted (14). It was subsequently determined that the Mnand Zn-binding S100 protein calprotectin (CP) is a critical component of this nutrient-withholding response (10,14,18,19). CPdeficient mice fail to remove Mn, but not Zn, from staphylococcal liver abscesses and have higher bacterial burdens during infection (14, 18).…”

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confidence: 99%

MntABC and MntH Contribute to Systemic Staphylococcus aureus Infection by Competing with Calprotectin for Nutrient Manganese

et al. 2013

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During infection, vertebrates limit access to manganese and zinc, starving invading pathogens, such as Staphylococcus aureus, of these essential metals in a process termed "nutritional immunity." The manganese and zinc binding protein calprotectin is a key component of the nutrient-withholding response, and mice lacking this protein do not sequester manganese from S. aureus liver abscesses. One potential mechanism utilized by S. aureus to minimize host-imposed manganese and zinc starvation is the expression of the metal transporters MntABC and MntH. We performed transcriptional analyses of both mntA and mntH, which revealed increased expression of both systems in response to calprotectin treatment. MntABC and MntH compete with calprotectin for manganese, which enables S. aureus growth and retention of manganese-dependent superoxide dismutase activity. Loss of MntABC and MntH results in reduced staphylococcal burdens in the livers of wild-type but not calprotectin-deficient mice, suggesting that these systems promote manganese acquisition during infection. During the course of these studies, we observed that metal content and the importance of calprotectin varies between murine organs, and infection leads to profound changes in the anatomical distribution of manganese and zinc. In total, these studies provide insight into the mechanisms utilized by bacteria to evade host-imposed nutrient metal starvation and the critical importance of restricting manganese availability during infection. Staphylococcus aureus is a commensal organism that asymptomatically colonizes nearly one-third of the population (1). However, once S. aureus breaches the epithelial barrier, the bacterium is capable of infecting nearly every organ despite the robust defenses elaborated by the host (2). This adaptability contributes to the significant morbidity and mortality associated with S. aureus infections. The emergence of methicillin-and vancomycin-resistant isolates has compounded the threat of this organism, highlighting the need to identify new therapeutics (3-7). This is of particular importance, as antibiotic resistance is prevalent in both hospital-and community-acquired isolates (4,6,8).Metals are essential for all forms of life due to their critical contributions to protein structure and enzymatic function (9-12). To combat invading pathogens, vertebrates leverage the essentiality of transition metals by restricting their availability, a process termed "nutritional immunity" (10, 13). While the most prominent example of nutritional immunity is the restriction of iron (Fe) by the host, it has recently been discovered that vertebrates also limit manganese (Mn) and zinc (Zn) availability during infection (10,(13)(14)(15)(16)(17). In fact, examination of abscesses formed during S. aureus infection has revealed that these lesions are Mn and Zn depleted (14). It was subsequently determined that the Mnand Zn-binding S100 protein calprotectin (CP) is a critical component of this nutrient-withholding response (10,14,18,19). CPdeficient mice...

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Human Calprotectin: Effect of Calcium and Zinc on its Secondary and Tertiary Structures, and Role of pH in its Thermal Stability

Cited by 34 publications

References 42 publications

Myeloperoxidase-dependent Lipid Peroxidation Promotes the Oxidative Modification of Cytosolic Proteins in Phagocytic Neutrophils

Myeloperoxidase-dependent Lipid Peroxidation Promotes the Oxidative Modification of Cytosolic Proteins in Phagocytic Neutrophils

Molecular basis for manganese sequestration by calprotectin and roles in the innate immune response to invading bacterial pathogens

MntABC and MntH Contribute to Systemic Staphylococcus aureus Infection by Competing with Calprotectin for Nutrient Manganese

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