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Sumin Lee

Publications and source records attributed to Sumin Lee.

2 recordsLinked to original sources

An Instrumental Optimization of a Label-Free Proteomic Method for Trace Protein Input.

Liquid chromatography-mass spectrometry (LC-MS)-based proteomics of trace-level samples, such as tens of cells or spatially resolved tissue regions, offers unique biological insights but is often constrained by the requirement for specialized, costly instrumentation. In this study, we developed a scalable workflow for the deep proteomic analysis of low- to ultralow-input samples by systematically optimizing a widely adopted Orbitrap and UHPLC platform to maximize sensitivity, precision, and throughput. This optimized workflow identified over 5600 proteins from 5 ng of peptides and 3400 proteins from 20 sorted cells, achieving a throughput of 30 analyses per day while maintaining deep proteome coverage and high quantitative reproducibility. Furthermore, by applying this method to spatially resolved proteomics, we identified over 6100 proteins from microscale regions of interest (ROIs) within a formalin-fixed, paraffin-embedded (FFPE) tissue. A data-driven normalization strategy was employed to correct for variable cellularity across tissue regions, effectively revealing intratumor heterogeneity and distinct molecular and functional signatures, including pathway activations not apparent in parallel spatial transcriptomic analysis. Ultimately, this accessible, high-performance method substantially lowers the instrumentation barrier for the deep proteomic profiling of trace-level biological samples.

Proteomics

Mutation of the gidB gene causes intrinsic streptomycin resistance in Bacillus velezensis.

Bacillus velezensis strain DMB07, isolated from the traditional fermented Korean soybean meju, exhibits resistance to streptomycin [minimum inhibitory concentration (MIC) 128 mg/L]. To shed light on the genetic background behind this phenotype, this study determined the complete genome sequence of strain DMB07 and compared it with the genomes of two B. velezensis strains that are sensitive to streptomycin. Compared with the streptomycin-sensitive strains, in strain DMB07 there was a mutation of a nucleotide (C58T) of the 16 S rRNA (guanine527-N7)-methyltransferase gene (gidB) that leads to a change in the amino acid sequence of the protein (Arg20Cys). This sequence of gidB gene was previously linked with streptomycin resistance. To test the hypothesis that this change in the gidB gene sequence of strain DMB07 confers streptomycin resistance, a temperature-sensitive plasmid, pIMAY-tgidBT58C, was constructed for site-directed mutation (from thymine to cytosine) of nucleotide 58 of gidB in strain DMB07. The resulting strain, DMB07gidBT58C, showed the decreased MIC value (32 mg/L) against streptomycin. Furthermore, introduction of the wild-type gidB gene into strain DMB07gidBT58C resulted in recovery of the MIC for streptomycin to 128 mg/L. Thus, a single mutation of the nucleotide sequence of the gidB gene can confer resistance to streptomycin.

Streptomycin