RT Journal Article
SR Electronic
T1 Structural Features and Oligomeric Nature of Human Podocin Domain
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.05.21.108415
DO 10.1101/2020.05.21.108415
A1 Sandeep KN Mulukala
A1 Shivkumar S Irukuvajjula
A1 Krishan Kumar
A1 Kanchan Garai
A1 Pannuru Venkatesu
A1 Ramakrishna Vadrevu
A1 Anil K Pasupulati
YR 2020
UL http://biorxiv.org/content/early/2020/05/22/2020.05.21.108415.abstract
AB Podocytes are crucial cells of the glomerular filtration unit and playing a vital role at the interface of the blood-urine barrier. Podocyte slit-diaphragm is a modified tight junction that facilitates size and shape-dependent permselectivity. Several proteins including podocin, nephrin, CD2AP, and TRPC6 form a macromolecular assembly and constitute the slit-diaphragm. Podocin is an integral membrane protein attached to the inner membrane of the podocyte via a short transmembrane region (101-125). The cytosolic N- and C-terminus help podocin to attain a hook-like structure. Podocin shares 44% homology with stomatin family proteins and similar to the stomatin proteins, podocin was shown to associate into higher-order oligomers at the site of slit-diaphragm. However, the stoichiometry of the homo-oligomers and how it partakes in the macromolecular assemblies with other slit-diaphragm proteins remains elusive. Here we investigated the oligomeric propensity of a truncated podocin construct (residues:126-350). We show that the podocin domain majorly homo-oligomerize into a 16mer. Circular dichroism and fluorescence spectroscopy suggest that the 16mer oligomer has considerable secondary structure and moderate tertiary packing.Competing Interest StatementThe authors have declared no competing interest.SDslit-diaphragmGFBGlomerular filtration barrierCD2APCD-2 associated proteinTRPC6Transient receptor potential cation channel subfamily C member 6ZO-1Zonula occludens-1NEPHNephrin-like proteinNSNephrotic syndromeSRNSsteroid-resistant NSNPHS1 & 2Nephrotic syndrome-type I and type IIIDRsIntrinsically disordered regionsSECSize-exclusion chromatographyMALSmulti-angle light scatteringCDCircular dichroism