Quantitative Differences in the Urinary Proteome of Siblings Discordant for Type 1 Diabetes Include Lysosomal Enzymes

Abstract

Individuals with type 1 diabetes (T1D) often have higher than normal blood glucose levels, causing advanced glycation end product formation and inflammation and increasing the risk of vascular complications years or decades later. To examine the urinary proteome in juveniles with T1D for signatures indicative of inflammatory consequences of hyperglycemia, we profiled the proteome of 40 T1D patients with an average of 6.3 years after disease onset and normal or elevated HbA1C levels, in comparison with a cohort of 41 healthy siblings. Using shotgun proteomics, 1036 proteins were identified, on average, per experiment, and 50 proteins showed significant abundance differences using a Wilcoxon signed-rank test (FDR q-value ≤ 0.05). Thirteen lysosomal proteins were increased in abundance in the T1D versus control cohort. Fifteen proteins with functional roles in vascular permeability and adhesion were quantitatively changed, including CD166 antigen and angiotensin-converting enzyme 2. α-N-Acetyl-galactosaminidase and α-fucosidase 2, two differentially abundant lysosomal enzymes, were detected in western blots with often elevated quantities in the T1D versus control cohort. Increased release of proteins derived from lysosomes and vascular epithelium into urine may result from hyperglycemia-associated inflammation in the kidney vasculature.

Case report: intermittent fasting and probiotic yogurt consumption are associated with reduction of serum alpha-N-acetylgalactosaminidase and increased urinary excretion of lipophilic toxicants

“…(15) It has been proposed that nagalase is a marker of cancer cell proliferation (16) as well as of systemic inflammation. (17) Therefore, the observed decrease of serum nagalase activity, together with the decrease of lymphocytes accompanied by an increase of platelets, may reflect a positive impact of the nutritional approach described herein on the underlying pathologic condition of Dr. JB. Such a positive impact may also be consistent with the observed decrease of serum TSH values.…”

Nutritional Immunity and Antibiotic Drug Treatments Influence Microbial Composition but Fail to Eliminate Urethral Catheter Biofilms in Recurrently Catheterized Patients

“…Both systems (Thermo Scientific, San Jose, CA) were equipped with a FLEX nano-electrospray ion source at the LC-MS interface. Analytic procedures were previously described for the Q-Exactive (69, 70) and LTQ-Velos Pro (71) platforms. For LTQ-Velos Pro analysis, peptides present in a sample were trapped on a C 18 trap column (100 μm × 2 cm, 5 μm pore size, 120 Å) and separated on a PicoFrit C 18 analytical column (75 μm × 15 cm, 3 μm pore size, 150 Å) at a flow rate of 200 nl/min.…”

<em>Actinobaculum massiliense</em> Proteome Profiled in Polymicrobial Urethral Catheter Biofilms

“…The LC-MS/MS workstation was composed of the LTQ-Velos Pro iontrap mass spectrometer coupled to the Easy-nLC II system via a FLEX nano-electrospray ion source (Thermo Scientific, San Jose, CA). Detailed LC-MS/MS analysis steps were previously described [14]. The sample was loaded onto a C18 trap column (100 μm × 2 cm, 5 μm pore size, 120 Å) and separated on a PicoFrit C 18 analytical column (75 μm × 15 cm, 3 μm pore size, 150 Å) at a flow rate of 200 nL/min.…”