enzyme fusion
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Metabolites ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 125
Author(s):  
Xueqin Lv ◽  
Shixiu Cui ◽  
Yang Gu ◽  
Jianghua Li ◽  
Guocheng Du ◽  
...  

Enzyme assembly by ligand binding or physically sequestrating enzymes, substrates, or metabolites into isolated compartments can bring key molecules closer to enhance the flux of a metabolic pathway. The emergence of enzyme assembly has provided both opportunities and challenges for metabolic engineering. At present, with the development of synthetic biology and systems biology, a variety of enzyme assembly strategies have been proposed, from the initial direct enzyme fusion to scaffold-free assembly, as well as artificial scaffolds, such as nucleic acid/protein scaffolds, and even some more complex physical compartments. These assembly strategies have been explored and applied to the synthesis of various important bio-based products, and have achieved different degrees of success. Despite some achievements, enzyme assembly, especially in vivo, still has many problems that have attracted significant attention from researchers. Here, we focus on some selected examples to review recent research on scaffold-free strategies, synthetic artificial scaffolds, and physical compartments for enzyme assembly or pathway sequestration, and we discuss their notable advances. In addition, the potential applications and challenges in the applications are highlighted.


2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Ruben J. Boado ◽  
Jeff Zhiqiang Lu ◽  
Eric Ka-Wai Hui ◽  
Huilan Lin ◽  
William M. Pardridge

AbstractMost lysosomal storage disorders affect the central nervous system. However, lysosomal enzymes do not cross the blood-brain barrier (BBB), and intravenous enzyme infusion is not effective for the brain. Lysosomal enzymes can be re-engineered for BBB transport as IgG-enzyme fusion proteins, where the IgG domain is a monoclonal antibody (MAb) against an endogenous BBB receptor/transporter, and which acts as a molecular Trojan horse to deliver the enzyme to brain. However, the problem is retention of high enzyme activity following enzyme fusion to the IgG. The present investigation shows this is possible with a versatile approach that employs fusion of the enzyme to either the IgG heavy chain or light chain using a long flexible linker. The model IgG is a chimeric monoclonal antibody (MAb) against the human insulin receptor (HIR). The enzyme activity of the HIRMAb-enzyme fusion protein is preserved for hexosaminidase A, which is mutated in Tay Sachs disease, for protein palmitoylthioesterase-1, which is mutated in Batten disease type 1, acid sphingomyelinase, which is mutated in Niemann Pick disease type A, and beta galactosidase-1, which is mutated in GM1 gangliosidosis.


2019 ◽  
Vol 30 (4) ◽  
pp. 689-705.e6 ◽  
Author(s):  
M. Kathryn Brewer ◽  
Annette Uittenbogaard ◽  
Grant L. Austin ◽  
Dyann M. Segvich ◽  
Anna DePaoli-Roach ◽  
...  

2019 ◽  
Vol 67 (35) ◽  
pp. 9851-9857 ◽  
Author(s):  
Hu-Hu Liu ◽  
Chong Wang ◽  
Xiang-Yang Lu ◽  
He Huang ◽  
Yun Tian ◽  
...  

2019 ◽  
Vol 16 (9) ◽  
pp. 3791-3801 ◽  
Author(s):  
Grant L. Austin ◽  
Zoe R. Simmons ◽  
Jack E. Klier ◽  
Alberto Rondon ◽  
Brad L. Hodges ◽  
...  

2019 ◽  
Author(s):  
M. Kathryn Brewer ◽  
Annette Uittenbogaard ◽  
Grant Austin ◽  
John J. McCarthy ◽  
Dyann M. Segvich ◽  
...  

AbstractLafora disease (LD) is a fatal childhood epilepsy and a non-classical glycogen storage disorder with no effective therapy or cure. LD is caused by recessive mutations in theEPM2AorEPM2Bgenes that encode the glycogen phosphatase laforin and an E3 ubiquitin ligase malin, respectively. A hallmark of LD is the intracellular accumulation of abnormal and insoluble α-linked polysaccharide deposits known as Lafora bodies (LBs) in several tissues, including most regions of the brain. In mouse models of LD, genetic reduction of glycogen synthesis eliminates LB formation and rescues the neurological phenotype. Since multiple groups have confirmed that neurodegeneration and epilepsy result from LB accumulation, a major focus in the field has shifted toward the development of therapies that reduce glycogen synthesis or target LBs for degradation with the goal of treating LD. Herein, we identify the optimal enzymes for degrading LBs, and we develop a novel therapeutic agent by fusing human pancreatic α-amylase to a cellpenetrating antibody fragment. This antibody-enzyme fusion (VAL-0417) degrades LBsin vitro, shows robust cellular uptake, and significantly reduces the LB loadin vivoinEpm2a-/- mice. VAL-0417 is a promising therapeutic for the treatment of LD and a putative precision therapy for an intractable epilepsy. Antibody-enzyme fusions represent a new class of antibody-based drugs that could be utilized to treat glycogen storage disorders and other diseases.One Sentence SummaryAn antibody-enzyme fusion delivering an amylase degrades the toxic polyglucosan bodies that cause Lafora disease, a fatal childhood epilepsy.


2019 ◽  
Author(s):  
Grant L. Austin ◽  
Zoe R. Simmons ◽  
Jack E. Klier ◽  
Brad L. Hodges ◽  
Robert Shaffer ◽  
...  

AbstractLafora disease is a fatal juvenile epilepsy, characterized by the malignant accumulation of aberrant glucan inclusions called Lafora Bodies (LBs). Cerebral delivery of protein-based therapeutics for the clearance of Lafora Bodies remain a unique challenge in the field. Recently, a humanized antigen-binding fragment (hFab) derived from a murine systemic lupus erythematosus DNA autoantibody (3E10) has been shown to mediate cell penetration and been proposed as a broadly applicable carrier to mediate cellular targeting and uptake. We report studies on cerebral delivery of VAL-0417, an antibody-enzyme fusion composed of the 3E10 hFab and human pancreatic α-amylase for the clearance of LBs in a mouse model of lafora disease. Herein, we report development of an enzyme-linked immunosorbant-based bioassay to detect VAL-0417 post treatment as a measure of delivery efficacy. We demonstrate the robust and sensitive detection of the fusion protein in multiple tissue types. Using our method, we measured biodistribution in different methods of delivery. We found intracerebroventricular administration provided the most robust delivery, while intrathecal administration only showed modest biodistribution. These data define critical steps in the translational pipeline of VAL-0417for the treatment of Lafora disease.


2019 ◽  
Vol 8 ◽  
pp. e00086 ◽  
Author(s):  
Hery Rabeharindranto ◽  
Sara Castaño-Cerezo ◽  
Thomas Lautier ◽  
Luis F. Garcia-Alles ◽  
Christian Treitz ◽  
...  

2019 ◽  
Vol 126 (2) ◽  
pp. S24
Author(s):  
Dustin Armstrong ◽  
Tracy McKnight ◽  
Nadine Aziz

2018 ◽  
Vol 179 (3) ◽  
pp. 1013-1027 ◽  
Author(s):  
Maurizio Camagna ◽  
Alexander Grundmann ◽  
Cornelia Bär ◽  
Julian Koschmieder ◽  
Peter Beyer ◽  
...  

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