Albert J de Neeling

Publications of Albert J de Neeling

• Synthesis and evaluation of strand and turn modified ring-extended gramicidin S derivatives.

  Authors: Annemiek D Knijnenburg, Varsha V Kapoerchan, Gijsbert M Grotenbreg, Emile Spalburg, Albert J de Neeling, Roos H Mars-Groenendijk, Daan Noort, José M Otero, Antonio L Llamas-Saiz, Mark J van Raaij, Bep Ravensbergen, Peter H Nibbering, Gijs A van der Marel, Herman S Overkleeft, Mark Overhand

  Bioorganic & medicinal chemistry. 06/2011; 19(11):3402-9.

  In this paper, we describe the crystal structure of previously reported ring-extended gramicidin S (GS) derivative 2 (GS14K4), containing a d-amino acid residue in one of the β-strand regions. ThisIn this paper, we describe the crystal structure of previously reported ring-extended gramicidin S (GS) derivative 2 (GS14K4), containing a d-amino acid residue in one of the β-strand regions. This structure is in agreement with a previously reported modeling study of the same molecule. The polar side chain of the additional d-amino acid residue is positioned at the same face of the molecule as the hydrophobic side chains, and we believe that because of this compound 2 is considerably less hydrophobic than extended GS derivatives in which the strand regions are exclusively composed of l-amino acids. Using this backbone structure as our benchmark we prepared a small series of ring-extended GS analogues featuring sugar amino acid dipeptide isosteres of varied hydrophobicity at the turn region. We show that via this approach hydrophobicity of extended GS analogues can be tuned without affecting the secondary structure (as observed from NMR and CD spectra). Biological evaluation reveals that hydrophobicity correlates to cell toxicity, but still bacteriolysis is induced with GS analogues that are too hydrophilic to efficiently lyse human red blood cells.

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• Exploring the conformational and biological versatility of β-turn-modified gramicidin S by using sugar amino acid homologues that vary in ring size.

  Authors: Annemiek D Knijnenburg, Adriaan W Tuin, Emile Spalburg, Albert J de Neeling, Roos H Mars-Groenendijk, Daan Noort, José M Otero, Antonio L Llamas-Saiz, Mark J van Raaij, Gijs A van der Marel, Herman S Overkleeft, Mark Overhand

  Chemistry (Weinheim an der Bergstrasse, Germany). 03/2011; 17(14):3995-4004.

  Monobenzylated sugar amino acids (SAAs) that differ in ether ring size (containing an oxetane, furanoid, and pyranoid ring) were synthesized and incorporated in one of the β-turn regions of theMonobenzylated sugar amino acids (SAAs) that differ in ether ring size (containing an oxetane, furanoid, and pyranoid ring) were synthesized and incorporated in one of the β-turn regions of the cyclo-decapeptide gramicidin S (GS). CD, NMR spectroscopy, modeling, and X-ray diffraction reveal that the ring size of the incorporated SAA moieties determines the spatial positioning of their cis-oriented carboxyl and aminomethyl substituents, thereby subtly influencing the amide linkages with the adjacent amino acids in the sequence. Unlike GS itself, the conformational behavior of the SAA-containing peptides is solvent dependent. The derivative containing the pyranoid SAA is slightly less hydrophobic and displays a diminished haemolytic activity, but has similar antimicrobial properties as GS.

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• Studies on Prn variation in the mouse model and comparison with epidemiological data.

  Authors: Marjolein van Gent, Inge H M van Loo, Kees J Heuvelman, Albert J de Neeling, Peter Teunis, Frits R Mooi

  PloS one. 01/2011; 6(3):e18014.

  The virulence factor pertactin (Prn) is a component of pertussis vaccines and one of the most polymorphic Bordetella pertussis antigens. After the introduction of vaccination shifts in predominantThe virulence factor pertactin (Prn) is a component of pertussis vaccines and one of the most polymorphic Bordetella pertussis antigens. After the introduction of vaccination shifts in predominant Prn types were observed and strains with the Prn vaccine type (Prn1) were replaced by strains carrying non-vaccine types (Prn2 and Prn3), suggesting vaccine-driven selection. The aim of this study was to elucidate the shifts observed in Prn variants. We show that, although Prn2 and Prn3 circulated in similar frequencies in the 1970s and 1980s, in the 1990s Prn2 strains expanded and Prn3 strains disappeared, suggesting that in vaccinated populations Prn2 strains are fitter than Prn3 strains. We established a role for Prn in the mouse model by showing that a Prn knock-out (Prn-ko) mutation reduced colonization in trachea and lungs. Restoration of the mutation resulted in a significant increase in colonization compared to the knock-out mutant. The ability of clinical isolates with different Prn variants to colonize the mouse lung was compared. Although these isolates were also polymorphic at other loci, only variation in the promoter for pertussis toxin (ptxP) and Prn were found to contribute significantly to differences in colonization. Analysis of a subset of strains with the same ptxP allele revealed that the ability to colonize mice decreased in the order Prn1>Prn2 and Prn3. Our results are consistent with the predominance of Prn1 strains in unvaccinated populations. Our results show that ability to colonize mice is practically the same for Prn2 and Prn3. Therefore other factors may have contributed to the predominance of Prn2 in vaccinated populations. The mouse model may be useful to assess and predict changes in the B. pertussis population due to vaccination.

• An adamantyl amino acid containing gramicidin S analogue with broad spectrum antibacterial activity and reduced hemolytic activity.

  Authors: Varsha V Kapoerchan, Annemiek D Knijnenburg, Miquel Niamat, Emile Spalburg, Albert J de Neeling, Peter H Nibbering, Roos H Mars-Groenendijk, Daan Noort, José M Otero, Antonio L Llamas-Saiz, Mark J van Raaij, Gijs A van der Marel, Herman S Overkleeft, Mark Overhand

  Chemistry (Weinheim an der Bergstrasse, Germany). 10/2010; 16(40):12174-81.

  The cyclic cationic antimicrobial peptide gramicidin S (GS) is an effective topical antibacterial agent that is toxic for human red blood cells (hemolysis). Herein, we present a series of amphiphilicThe cyclic cationic antimicrobial peptide gramicidin S (GS) is an effective topical antibacterial agent that is toxic for human red blood cells (hemolysis). Herein, we present a series of amphiphilic derivatives of GS with either two or four positive charges and characteristics ranging between very polar and very hydrophobic. Screening of this series of peptide derivatives identified a compound that combines effective antibacterial activity with virtually no toxicity within the same concentration range. This peptide acts against both Gram-negative and Gram-positive bacteria, including several MRSA strains, and represents an interesting lead for the development of a broadly applicable antibiotic.

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• Tuning hydrophobicity of highly cationic tetradecameric Gramicidin S analogues using adamantane amino acids.

  Authors: Annemiek D Knijnenburg, Varsha V Kapoerchan, Emile Spalburg, Albert J de Neeling, Roos H Mars-Groenendijk, Daan Noort, Gijs A van der Marel, Herman S Overkleeft, Mark Overhand

  Bioorganic & medicinal chemistry. 09/2010; 18(23):8403-9.

  Ring extended Gramicidin S analogues containing adamantane amino acids and six cationic residues were designed and evaluated. Systematic replacement of the hydrophobic residues with adamantane aminoRing extended Gramicidin S analogues containing adamantane amino acids and six cationic residues were designed and evaluated. Systematic replacement of the hydrophobic residues with adamantane amino acids resulted in a small set of compounds with varying amphipathic character. It was found that the amphipathicity of these compounds is correlated to their biological activity. Several bacterial strains including MRSA strains were shown to be killed by the novel peptides. The most potent antibacterial peptides are tetradecameric GS analogues containing six positives charges and two adamantane moieties.

• Gramicidin S derivatives containing cis- and trans-morpholine amino acids (MAAs) as turn mimetics.

  Authors: Varsha V Kapoerchan, Emile Spalburg, Albert J de Neeling, Roos H Mars-Groenendijk, Daan Noort, José M Otero, Patricia Ferraces-Casais, Antonio L Llamas-Saiz, Mark J van Raaij, Joop van Doorn, Gijs A van der Marel, Herman S Overkleeft, Mark Overhand

  Chemistry (Weinheim an der Bergstrasse, Germany). 03/2010; 16(14):4259-65.

  The cyclic decapeptide gramicidin S (GS) was used as a model for the evaluation of four turn mimetics. For this purpose, one of the D-Phe-Pro two-residue turn motifs in the rigid cyclic beta-hairpinThe cyclic decapeptide gramicidin S (GS) was used as a model for the evaluation of four turn mimetics. For this purpose, one of the D-Phe-Pro two-residue turn motifs in the rigid cyclic beta-hairpin structure of GS was replaced with morpholine amino acids (MAA 2-5), differing in stereochemistry and length of the side-chain. The conformational properties of the thus obtained GS analogues (6-9) was assessed by using NMR spectroscopy and X-ray crystallography, and correlated with their biological properties (antimicrobial and hemolytic activity). We show that compound 8, containing the dipeptide isostere trans-MAA 4, has an apparent high structural resemblance with GS and that its antibacterial activity against a panel of Gram positive and -negative bacterial strains is better than the derivatives 6, 7 and 9.

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• PFGE diversity within the methicillin-resistant Staphylococcus aureus clonal lineage ST398.

  Authors: Thijs Bosch, Albert J de Neeling, Leo M Schouls, Kim W van der Zwaluw, Jan A J W Kluytmans, Hajo Grundmann, Xander W Huijsdens

  BMC microbiology. 02/2010; 10:40.

  Livestock has recently been identified as a new reservoir of methicillin-resistant Staphylococcus aureus (MRSA). Most isolates belong to ST398 and are non-typeable with PFGE using SmaI, making itLivestock has recently been identified as a new reservoir of methicillin-resistant Staphylococcus aureus (MRSA). Most isolates belong to ST398 and are non-typeable with PFGE using SmaI, making it difficult to study transmission and outbreaks. Therefore, a new PFGE using Cfr9I, a neoschizomer of SmaI was optimized and evaluated to investigate ST398 isolates. After optimizing and evaluating the Cfr9I PFGE, clear and reproducible banding patterns were obtained from all previously non-typeable MRSA (NT(SmaI) -MRSA) isolates. The PFGE patterns of ST398 isolates showed more diversity than with spa-typing and/or MLST. The PFGE results showed diversity within and between the two most prevalent spa-types of NT(SmaI) -MRSA (t011 and t108). No match was found, when comparing banding patterns of the NT(SmaI) -MRSA with 700 different PFGE types, obtained with SmaI digestion, in our database of more than 4000 strains. Furthermore, possible transmission among veterinarians and their family members was investigated and an outbreak of ST398 MRSA in a residential care facility was confirmed with the Cfr9I PFGE. The adjusted PFGE can be used as a method for selecting important and distinct ST398 isolates for further research. The adjustments in the PFGE protocol using Cfr9I are easy to implement to study the ST398 clonal lineage in laboratories which already have a PFGE facility.

• Ring-Extended Derivatives of Gramicidin S with Furanoid Sugar Amino Acids in the Turn Region Have Enhanced Antimicrobial Activity.

  Authors: Annemiek D Knijnenburg, Emile Spalburg, Albert J de Neeling, Roos H Mars-Groenendijk, Daan Noort, Gijsbert M Grotenbreg, Gijs A van der Marel, Herman S Overkleeft, Mark Overhand

  ChemMedChem. 10/2009;

• Structural and biological evaluation of some loloatin C analogues.

  Authors: Adriaan W Tuin, Gijsbert M Grotenbreg, Emile Spalburg, Albert J de Neeling, Roos H Mars-Groenendijk, Gijsbert A van der Marel, Daan Noort, Herman S Overkleeft, Mark Overhand

  Bioorganic & medicinal chemistry. 08/2009;

  Loloatin C is a cyclic cationic antimicrobial peptide which is active against Gram positive as well as certain Gram negative bacteria. Unfortunately, it is equally potent against human erythrocytes.Loloatin C is a cyclic cationic antimicrobial peptide which is active against Gram positive as well as certain Gram negative bacteria. Unfortunately, it is equally potent against human erythrocytes. To probe the structure-activity relationship of this promising antibiotic peptide, amino acid substitution and/or incorporation of a constraint sugar amino acid dipeptide isoster has been applied. Six new derivatives have been synthesized using SPPS and their solution structure investigated using NMR studies. Finally, the antimicrobial and the hemolytic activities have been determined.

• Multiple-locus variable number tandem repeat analysis of Staphylococcus aureus: comparison with pulsed-field gel electrophoresis and spa-typing.

  Authors: Leo M Schouls, Emile C Spalburg, Martijn van Luit, Xander W Huijsdens, Gerlinde N Pluister, Marga G van Santen-Verheuvel, Han G J van der Heide, Hajo Grundmann, Max E O C Heck, Albert J de Neeling

  PLoS ONE. 02/2009; 4(4):e5082.

  BACKGROUND: Molecular typing of methicillin-resistant Staphylococcus aureus (MRSA) is required to study the routes and rates of transmission of this pathogen. Currently available typing techniquesBACKGROUND: Molecular typing of methicillin-resistant Staphylococcus aureus (MRSA) is required to study the routes and rates of transmission of this pathogen. Currently available typing techniques are either resource-intensive or have limited discriminatory ability. Multiple-locus variable number tandem repeat analysis (MLVA) may provide an alternative high throughput molecular typing tool with high epidemiological resolution. METHODOLOGY/PRINCIPAL FINDINGS: A new MLVA scheme for S. aureus was validated using 1681 S. aureus isolates collected from Dutch patients and 100 isolates from pigs. MLVA using 8 tandem repeat loci was performed in 2 multiplex PCRs and the fluorescently labeled PCR products were accurately sized on an automated DNA sequencer. The assessed number of repeats was used to create MLVA profiles consisting of strings of 8 integers that were used for categorical clustering. MLVA yielded 511 types that clustered into 11 distinct MLVA complexes which appeared to coincide with MLST clonal complexes. MLVA was at least as discriminatory as PFGE and twice as discriminatory as spa-sequence typing. There was considerable congruence between MLVA, spa-sequence typing and PFGE, at the MLVA complex level with group separation values of 95.1% and 89.2%. MLVA could not discriminate between pig-related MRSA strains isolated from humans and pigs, corroborating the high degree of relationship. MLVA was also superior in the grouping of MRSA isolates previously assigned to temporal-spatial clusters with indistinguishable SpaTypes, demonstrating its enhanced epidemiological usefulness. CONCLUSIONS: The MLVA described in this study is a high throughput, relatively low cost genotyping method for S. aureus that yields discrete and unambiguous data that can be used to assign biological meaningful genotypes and complexes and can be used for interlaboratory comparisons in network accessible databases. Results suggest that MLVA offsets the disadvantages of other high discriminatory typing approaches and represents a promising tool for hospital, national and international molecular epidemiology.

• Methicillin-resistant and -susceptible Staphylococcus aureus sequence type 398 in pigs and humans.

  Authors: Alex van Belkum, Damian C Melles, Justine K Peeters, Willem B van Leeuwen, Engeline van Duijkeren, Xander W Huijsdens, Emile Spalburg, Albert J de Neeling, Henri A Verbrugh, Dutch Working Party on Surveillance and Research of MRSA-SOM

  Emerging infectious diseases. 04/2008; 14(3):479-83.

  Methicillin-resistant Staphylococcus aureus sequence type 398 (ST398 MRSA) was identified in Dutch pigs and pig farmers. ST398 methicillin-susceptible S. aureus circulates among humans at lowMethicillin-resistant Staphylococcus aureus sequence type 398 (ST398 MRSA) was identified in Dutch pigs and pig farmers. ST398 methicillin-susceptible S. aureus circulates among humans at low frequency (0.2%) but was isolated in 3 human cases of bacteremia (2.1%; p = 0.026). Although its natural host is probably porcine, ST398 MRSA likely causes infections in humans.

• Beta-turn modified gramicidin S analogues containing arylated sugar amino acids display antimicrobial and hemolytic activity comparable to the natural product.

  Authors: Gijsbert M Grotenbreg, Annelies E M Buizert, Antonio L Llamas-Saiz, Emile Spalburg, Peter A V van Hooft, Albert J de Neeling, Daan Noort, Mark J van Raaij, Gijsbert A van der Marel, Herman S Overkleeft, Mark Overhand

  Journal of the American Chemical Society. 07/2006; 128(23):7559-65.

  This paper describes the design and synthesis of gramicidin S (GS) analogues 10a-c containing arylated sugar amino acids (SAAs) as a replacement of one of the two (D)Phe-Pro beta-turn regions. TheThis paper describes the design and synthesis of gramicidin S (GS) analogues 10a-c containing arylated sugar amino acids (SAAs) as a replacement of one of the two (D)Phe-Pro beta-turn regions. The cyclic, amphiphilic peptides adopt a beta-sheet conformation featuring an unusual reverse turn induced by the SAAs. The altered turn region induces a slight distortion of the antiparallel beta-sheet, as compared to GS; the overall geometry however closely resembles that of the nonarylated GS analogue 1. GS analogues 10a-c proved to be as active as the parent GS itself as antibacterial agents and are equally efficient in lysing red blood cells. These properties are in sharp contrast to the diminished biological activity displayed by 1. We conclude that the presence of aromaticity in the turn regions of GS derivatives is required for biological activity, whereas the native conformation of the beta-hairpin is not. Our findings may guide future research toward efficient and nonhemolytic GS analogues for combating bacterial infections.

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• Community-acquired MRSA and pig-farming.

  Authors: Xander W Huijsdens, Beatrix J van Dijke, Emile Spalburg, Marga G van Santen-Verheuvel, Max E O C Heck, Gerlinde N Pluister, Andreas Voss, Wim J B Wannet, Albert J de Neeling

  Annals of clinical microbiology and antimicrobials. 02/2006; 5:26.

  BACKGROUND: Sporadic cases of CA-MRSA in persons without risk-factors for MRSA carriage are increasing. CASE PRESENTATION: We report a MRSA cluster among family members of a pig-farmer, hisBACKGROUND: Sporadic cases of CA-MRSA in persons without risk-factors for MRSA carriage are increasing. CASE PRESENTATION: We report a MRSA cluster among family members of a pig-farmer, his co-workers and his pigs. Initially a young mother was seen with mastitis due to MRSA. Six months later her baby daughter was admitted to the hospital with pneumococcal otitis. After staying five days in hospital, the baby was found to be MRSA positive. At that point it was decided to look for a possible source, such as other family members and house-hold animals, including pigs on the farm, since those were reported as a possible source of MRSA earlier. Swabs were taken from the throat and nares of family members and co-workers. A veterinarian obtained swabs from the nares, throat and perineum of 10 pigs. Swabs were cultured following a national protocol to detect MRSA that included the use of an enrichment broth. Animal and human strains were characterized by PFGE, spa-typing, MLST analysis, SSCmec, AGR typing, and the detection for PVL, LukM, and TSST toxin genes. Three family members, three co-workers, and 8 of the 10 pigs were MRSA positive. With the exception of the initial case (the mother) all persons were solely colonized, with no signs of clinical infections. After digestion with SmaI, none of the strains showed any bands using PFGE. All isolates belonged to spa type t108 and ST398. CONCLUSION: 1. This report clearly shows clonal spread and transmission between humans and pigs in the Netherlands. 2. MLST sequence type 398 might be of international importance as pig-MRSA, since this type was shown earlier to be present in epidemiologically unrelated French pigs and pig-farmers. 3. Research is needed to evaluate whether this is a local problem or a new source of MRSA, that puts the until now successful Search and Destroy policy of the Netherlands at risk.

• Synthesis and biological evaluation of gramicidin S dimers.

  Authors: Gijsbert M Grotenbreg, Martin D Witte, Peter A V van Hoof, Emile Spalburg, Philipp Reiss, Daan Noort, Albert J de Neeling, Ulrich Koert, Gijsbert A van der Marel, Herman S Overkleeft, Mark Overhand

  Organic & biomolecular chemistry. 02/2005; 3(2):233-8.

  The design and synthesis of analogues of the cyclic beta-sheet gramicidin S (GS), having additional functionalities in their turn regions, is reported. The monomeric GS analogues were transformedThe design and synthesis of analogues of the cyclic beta-sheet gramicidin S (GS), having additional functionalities in their turn regions, is reported. The monomeric GS analogues were transformed into dimers and their activities towards biological membranes, through antimicriobial and hemolytic assays, were evaluated. Finally, conductivity measurements have been performed to elucidate ion channel forming properties.

• A practical synthesis of gramicidin s and sugar amino Acid containing analogues.

  Authors: Gijsbert M Grotenbreg, Martijn Kronemeijer, Mattie S M Timmer, Farid El Oualid, Renate M van Well, Martijn Verdoes, Emile Spalburg, Peter A V van Hooft, Albert J de Neeling, Daan Noort, Jacques H van Boom, Gijsbert A van der Marel, Herman S Overkleeft, Mark Overhand

  The Journal of organic chemistry. 12/2004; 69(23):7851-9.

  A practical gram-scale and high-yielding synthesis of the antimicrobial peptide gramicidin S is presented. An Fmoc-based solid-phase peptide synthesis protocol is employed for the generation of theA practical gram-scale and high-yielding synthesis of the antimicrobial peptide gramicidin S is presented. An Fmoc-based solid-phase peptide synthesis protocol is employed for the generation of the linear decapeptide precursor, which is cyclized in solution to afford the target compound. The versatility of our method is demonstrated by the construction of eight gramicidin S analogues (15a-h) having nonproteinogenic sugar amino acid residues (4-7) incorporated in the turn regions.

• Epidemic and nonepidemic multidrug-resistant Enterococcus faecium.

  Authors: Helen L Leavis, Rob J L Willems, Janetta Top, Emile Spalburg, Ellen M Mascini, Ad C Fluit, Andy Hoepelman, Albert J de Neeling, Marc J M Bonten

  Emerging infectious diseases. 10/2003; 9(9):1108-15.

  The epidemiology of vancomycin-resistant Entero- coccus faecium (VREF) in Europe is characterized by a large community reservoir. In contrast, nosocomial outbreaks and infections (without a communityThe epidemiology of vancomycin-resistant Entero- coccus faecium (VREF) in Europe is characterized by a large community reservoir. In contrast, nosocomial outbreaks and infections (without a community reservoir) characterize VREF in the United States. Previous studies demonstrated host-specific genogroups and a distinct genetic lineage of VREF associated with hospital outbreaks, characterized by the variant esp-gene and a specific allele-type of the purK housekeeping gene (purK1). We investigated the genetic relatedness of vanA VREF (n=108) and vancomycin-susceptible E. faecium (VSEF) (n=92) from different epidemiologic sources by genotyping, susceptibility testing for ampicillin, sequencing of purK1, and testing for presence of esp. Clusters of VSEF fit well into previously described VREF genogroups, and strong associations were found between VSEF and VREF isolates with resistance to ampicillin, presence of esp, and purK1. Genotypes characterized by presence of esp, purK1, and ampicillin resistance were most frequent among outbreak-associated isolates and almost absent among community surveillance isolates. Vancomycin-resistance was not specifically linked to genogroups. VREF and VSEF from different epidemiologic sources are genetically related; evidence exists for nosocomial selection of a subtype of E. faecium, which has acquired vancomycin-resistance through horizontal transfer.

• Synthesis and biological evaluation of novel turn-modified gramicidin S analogues.

  Authors: Gijsbert M Grotenbreg, Emile Spalburg, Albert J de Neeling, Gijsbert A van der Marel, Herman S Overkleeft, Jacques H van Boom, Mark Overhand

  Bioorganic & medicinal chemistry. 08/2003; 11(13):2835-41.

  The synthesis of novel gramicidin S analogues having additional functionalities in the turn region by employing a biomimetic approach is described. The preservation of beta-sheet character in allThe synthesis of novel gramicidin S analogues having additional functionalities in the turn region by employing a biomimetic approach is described. The preservation of beta-sheet character in all analogues was established by NMR and the biological activity was evaluated.