Generation of phosphorylated proteins for structural studies

Structural studies of phosphorylated high mannose-type oligosaccharides.

Phosphomannosyl residues on acid hydrolases serve as recognition markers which target these enzymes to lysosomes. We have found that the oligosaccharide units of newly synthesized beta-glucuronidase contain phosphate residues in diester linkage between mannose and alpha-linked N-acetylglucosamine residues (Tabas, I., and Kornfeld, S. (1980) J. Biol. Chem. 255, 6633--6639). To obtain larger amou...

Structural Studies on Cytoskeletal Proteins.

A number of componenets of cytoskeleton are very large proteins which are in their complete native form non amenable to the structure determination via X-ray diffraction techniques due to their non-globular shape and intrinsic flexibility which hinder the formation of suitable crystals. Dissecting such proteins into their representative building blocks-domains and subsequent structural studies ...

Structural studies on lysosomal proteins

Structural and mechanistic studies of VPS4 proteins.

VPS4 ATPases function in multivesicular body formation and in HIV-1 budding. Here, we report the crystal structure of monomeric apo human VPS4B/SKD1 (hVPS4B), which is composed of five distinct elements: a poorly ordered N-terminal MIT domain that binds ESCRT-III substrates, large (mixed alpha/beta) and small (alpha) AAA ATPase domains that closely resemble analogous domains in the p97 D1 ATPas...

Structural Studies of Sugar Binding Proteins

Binding proteins, which are themselves non-enzymatic, play an important role in enzymatic reactions as well as non-enzymatic processes by providing a binding platform for the specific recognition of particular molecules. For example, periplasmic binding proteins play a vital role in nutrient uptake in Gram-negative bacteria. In the present study, three sugar binding proteins, including two peri...