Molecular organisation of the ice nucleation protein InaV from <i>Pseudomonas syringae</i>

Schmid, Daniel; Pridmore, David; Capitani, Guido; Battistutta, Roberto; Neeser, Jean‐Richard; Jann, Alfred

doi:10.1016/s0014-5793(97)01079-x

Cited by 88 publications

(85 citation statements)

References 22 publications

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“…Protein function is controlled by secondary and tertiary structures and in IN bacteria also by the formation of complexes (e.g. Kozloff et al, 1991;Schmid et al, 1997;Garnham et al, 2011). Heat is a simple but effective tool to disrupt and denature these proteinaceous INP.…”

Section: Heat Treatmentsmentioning

confidence: 99%

Sources of organic ice nucleating particles in soils

et al. 2016

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Abstract. Soil organic matter (SOM) may be a significant source of atmospheric ice nucleating particles (INPs), especially of those active > −15 • C. However, due to both a lack of investigations and the complexity of the SOM itself, the identities of these INPs remain unknown. To more comprehensively characterize organic INPs we tested locally representative soils in Wyoming and Colorado for total organic By contrast, lysozyme, which digests bacterial cell walls, only reduced INPs active at ≥ −7.5 or ≥ −6 • C, depending on the soil. The known IN bacteria were not detected in any soil, using PCR for the ina gene that codes for the active protein. We directly isolated and photographed two INPs from soil, using repeated cycles of freeze testing and subdivision of droplets of dilute soil suspensions; they were complex and apparently organic entities. Ice nucleation activity was not affected by digestion of Proteinase K-susceptible proteins or the removal of entities composed of fulvic and humic acids, sterols, or aliphatic alcohol monolayers. Organic INPs active colder than −10 to −12 • C were resistant to all investigations other than heat, oxidation with H 2 O 2 , and, for some, digestion with papain. They may originate from decomposing plant material, microbial biomass, and/or the humin component of the SOM. In the case of the latter then they are most likely to be a carbohydrate. Reflecting the diversity of the SOM itself, soil INPs have a range of sources which occur with differing relative abundances.

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Section: Heat Treatmentsmentioning

confidence: 99%

Sources of organic ice nucleating particles in soils

et al. 2016

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“…It is concluded that oligomers consisting of two up to a few single proteins could correspond to class III IN, i.e., initiate freezing in the temperature range from about −7 • C to −10 • C (Govindarajan and Lindow, 1988;Garnham et al, 2011), where the majority of the INA bacteria can be ice nucleation active. Larger protein oligomers are considered to be active at higher temperatures and are thought to be related to class I and II bacterial IN (Schmid et al, 1997). Up to now, the structure and functionality of the INA protein oligomers has not been finally clarified and is still the object of contemporary research (Garnham et al, 2011).…”

Section: S Hartmann Et Al: Immersion Freezing Of Ina Protein Complementioning

confidence: 99%

“…The INA proteins have been repeatedly shown to aggregate in bacterial outer membranes (e.g., Govindarajan and Lindow, 1988;Southworth et al, 1988;Mueller et al, 1990;Schmid et al, 1997). Govindarajan and Lindow (1988) estimated that two to more than a hundred proteins can be found in such an aggregate.…”

Section: Introductionmentioning

confidence: 99%

Immersion freezing of ice nucleation active protein complexes

et al. 2013

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Utilising the Leipzig Aerosol Cloud Interaction Simulator (LACIS), the immersion freezing behaviour of droplet ensembles containing monodisperse particles, generated from a Snomax™ solution/suspension, was investigated. Thereto ice fractions were measured in the temperature range between −5 °C to −38 °C. Snomax™ is an industrial product applied for artificial snow production and contains Pseudomonas syringae} bacteria which have long been used as model organism for atmospheric relevant ice nucleation active (INA) bacteria. The ice nucleation activity of such bacteria is controlled by INA protein complexes in their outer membrane.

In our experiments, ice fractions increased steeply in the temperature range from about −6 °C to about −10 °C and then levelled off at ice fractions smaller than one. The plateau implies that not all examined droplets contained an INA protein complex. Assuming the INA protein complexes to be Poisson distributed over the investigated droplet populations, we developed the CHESS model (stoCHastic modEl of similar and poiSSon distributed ice nuclei) which allows for the calculation of ice fractions as function of temperature and time for a given nucleation rate. Matching calculated and measured ice fractions, we determined and parameterised the nucleation rate of INA protein complexes exhibiting class III ice nucleation behaviour. Utilising the CHESS model, together with the determined nucleation rate, we compared predictions from the model to experimental data from the literature and found good agreement.

We found that (a) the heterogeneous ice nucleation rate expression quantifying the ice nucleation behaviour of the INA protein complex is capable of describing the ice nucleation behaviour observed in various experiments for both, Snomax™ and P. syringae bacteria, (b) the ice nucleation rate, and its temperature dependence, seem to be very similar regardless of whether the INA protein complexes inducing ice nucleation are attached to the outer membrane of intact bacteria or membrane fragments, (c) the temperature range in which heterogeneous droplet freezing occurs, and the fraction of droplets being able to freeze, both depend on the actual number of INA protein complexes present in the droplet ensemble, and (d) possible artifacts suspected to occur in connection with the drop freezing method, i.e., the method frequently used by biologist for quantifying ice nucleation behaviour, are of minor importance, at least for substances such as P. syringae, which induce freezing at comparably high temperatures. The last statement implies that for single ice nucleation entities such as INA protein complexes, it is the number of entities present in the droplet population, and the entities' nucleation rate, which control the freezing behaviour of the droplet population. Quantities such as ice active surface site density are not suitable in this context.

The results obtained in this study allow a different perspective on the quantification of the immersi...

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“…1988; Schmid et al, 1997;Southworth et al, 1988). These low-molecular-weight bands may represent degradation products of Ina proteins before and during preparation.…”

Section: Discussionmentioning

confidence: 99%

“…Ice nucleation proteins from various bacterial species have related internally repetitive primary structures (Michigami et al, 1995a;Schmid et al, 1997;Warren and Wolber, 1991). Each protein has three domains: a central repeating domain of 80% total sequence with an 8-amino acid residue period of repetition, a unique N-terminal domain, and a unique C-terminal domain.…”

Section: Discussionmentioning

confidence: 99%

Enhanced production of extracellular ice nucleators from Erwinia herbicola.

Lee

1998

J. Gen. Appl. Microbiol.

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Molecular organisation of the ice nucleation protein InaV from Pseudomonas syringae

Cited by 88 publications

References 22 publications

Sources of organic ice nucleating particles in soils

Sources of organic ice nucleating particles in soils

Immersion freezing of ice nucleation active protein complexes

Enhanced production of extracellular ice nucleators from Erwinia herbicola.

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