Dataset Information

Differentially expressed extracellular proteins of Leptospira under the physiological conditions of infection

ABSTRACT: Leptospirosis is a re-emerging zoonotic disease of global presence. If not diagnosed early, the disease progress to multi-organ failure and culminate in death. There is no effective vaccine or unambiguous early diagnostic methods to date. Extracellular proteins are expressed under pathogenic and invasion conditions will be useful targets for diagnostic measures. Leptospires comes to the water environment through the urine of reservoir animal hosts and stays long to infect human who gets in contact with the contaminated water. During infection two major physiological changes have to be encountered by the leptospires. One is the temperature that about 22 - 27 ºC of environmental water to humans at 37 °C and secondly, the osmolarity of environmental water 66 mOsm to 300 mOsm of humans. We have challenged mid-log phase leptospires to 37 ºC, 300 mOsm, and 37 °C + 300 mOsm and standard culture conditions as a control in EMJH medium for 24 h. The leptospires were harvested by centrifugation at 2,500 × g for 30 min at 4 °C. The Leptospira pellet was washed with PBS containing 5 mM MgCl2 three times were pooled and named as a “wash” for proteomic analysis (LC-MS/MS). The culture supernatant was centrifuged second time at 6000 × g for 30 min and a third time at 12,000 × g for 30 min to remove any Leptospira left in the medium, without tight packing of the cells and lysis, and is used to enrich non-abundant proteins using ProteoMiner™ Protein Enrichment Kit (Bio-Rad) and the enriched proteins were named “supernatant” and used for LC-MS/MS analysis. The samples from Standard culture, 37 ºC, 300 mOsm, and 37 °C + 300 mOsm were tagged with Tandem Mass Tag (TMT) (Thermo Fisher Scientific) followed by LCMS/MS analysis. The analysis showed differential expression of many proteins in wash and supernatant with respect to the standard culture condition.

INSTRUMENT(S):

ORGANISM(S): Leptospira Interrogans Serovar Copenhageni Str. Fiocruz L1-130 Leptospira

SUBMITTER: Madathiparambil G. Madanan

LAB HEAD: Dr Madathiparambil G. Madanan

PROVIDER: PXD037702 | Pride | 2025-09-08

REPOSITORIES: Pride

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Publications

Exoproteome Profiling Reveals Increased Secretion of Adhesins and Proteases by to Facilitate Host Colonization and Immune Modulation.

Sarma Abhijit A Phukan Homen H Devasahayam Arokia Balaya Rex R Thoduvayil Sikha S Dhandapani Gunasekaran G Christie Swamidurai Arul Diana SAD Prasad Thottethodi Subrahmanya Keshava TSK Madanan Madathiparambil Gopalakrishnan MG

ACS omega 20250727 30

Leptospirosis, a re-emerging zoonotic disease, is challenging human and animal health due to the lack of early and rapid diagnostic tools and effective vaccines. The exoproteome of the pathogen expressed under pathogenic conditions possesses a rational diagnostic significance due to its consistent presence in body fluids. were challenged to conditions simulating infection using physiological temperatures and osmolarity. Using state-of-the-art extraction techniques, efficient enrichment of nonabu ...[more]

PMID: 40787326

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Project description:Leptospirosis, an emerging zoonotic disease with worldwide distribution, is caused by spirochetes belonging to the genus Leptospira. More than 500,000 cases of severe leptospirosis are reported annually, with .10% of these being fatal. Leptospires can survive for weeks in suitably moist conditions before encountering a new host. Reservoir hosts, typically rodents, exhibit little to no signs of disease but shed large numbers of organisms in their urine. Transmission occurs when mucosal surfaces or abraded skin come into contact with infected urine or urine-contaminated water or soil. In humans, leptospires can cause a variety of clinical manifestations, ranging from asymptomatic or mild fever to severe icteric (Weil’s) disease and pulmonary haemorrhage. Currently, little is known about how Leptospira persist within a reservoir host. Prior in vitro studies have suggested that leptospires alter their transcriptomic and proteomic profiles in response to environmental signals encountered during mammalian infection. However, no study has examined gene expression by leptospires within a mammalian host-adapted state. To obtain a more faithful representation of how leptospires respond to host-derived signals, we used RNA-Seq to compare the transcriptome of L. interrogans cultivated within dialysis membrane chambers (DMCs) implanted into the peritoneal cavities of rats with that of organisms grown in vitro. In addition to determining the relative expression levels of ‘‘core’’ housekeeping genes under both growth conditions, we identified 166 genes that are differentially-expressed by L. interrogans in vivo. Our analyses highlight physiological aspects of host adaptation by leptospires relating to heme uptake and utilization. We also identified 11 novel non-coding transcripts that are candidate small regulatory RNAs. The DMC model provides a facile system for studying the transcriptional and antigenic changes associated with mammalian host-adaption, selection of targets for mutagenesis, and the identification of previously unrecognized virulence determinants. Transcriptome analysis of L. interrogans Copenhageni FIOCRUZ L1-130 using RNA from 2 different conditions using RNA-seq. Also, the reproducibility and robustness of data is ensured by three biological replicates from each condition.

			Action	DRS
	Lept_S-_4_.mzML	Mzml
	Lept_S-_4_.pep.xml	Pepxml
	Lept_SP_SCX3_R1.raw	Raw
	Lept_SP_SCX3_R2.raw	Raw
	Lept_SP_SCX3_R3.raw	Raw