Covalently bonded and adventitious glycans in germin.

Jaikaran, Anna S. I.; Kennedy, Theresa D.; Dratewka-Kos, Ella; Lane, B. G.

doi:10.1016/s0021-9258(19)38374-7

Cited by 69 publications

(5 citation statements)

References 57 publications

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“…They play an important role against diverse environmental stresses. Initially, GER was identified in wheat embryos as a germination-specific marker, but later, they were classified as a homohexamer glycoprotein with oxalate oxidase (OXO) activity, , while proteins showing an average similarity of 50% with it were called germin-like protein (GLPs). GLPs play an important role in diverse plant processes via oxalate oxidase (OXO), superoxide dismutase (SOD), glucophosphodiesterase, and polyphenol oxidase (PPO) enzymatic activities.…”

Section: Introductionmentioning

confidence: 99%

Molecular Characterization of Germin-like Protein Genes in Zea mays (ZmGLPs) Using Various In Silico Approaches

Ilyas¹,

Ali

Binjawhar

et al. 2023

ACS Omega

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Germin (GER) and germin-like proteins (GLPs) play an important role in various plant processes. Zea mays contains 26 germin-like protein genes ( ZmGLPs ) located on chromosomes 2, 4, and 10; most of which are functionally unexplored. The present study aimed to characterize all ZmGLPs using the latest computational tools. All of them were studied at a physicochemical, subcellular, structural, and functional level, and their expression was predicted in plant development, against biotic and abiotic stresses using various in silico approaches. Overall, ZmGLPs showed greater similarity in their physicochemical properties, domain architecture, and structure, mostly localized in the cytoplasmic or extracellular regions. Phylogenetically, they have a narrow genetic background with a recent history of gene duplication events on chromosome 4. Functional analysis revealed novel enzymatic activities of phosphoglycolate phosphatase, adenosylhomocysteinase, phosphoglycolate phosphatase-like, osmotin/thaumatin-like, and acetohydroxy acid isomeroreductase largely mediated by disulfide bonding. Expression analysis revealed their crucial role in the root, root tips, crown root, elongation and maturation zones, radicle, and cortex with the highest expression being observed during germination and at the maturity levels. Further, ZmGLPs showed strong expression against biotic ( Aspergillus flavus , Colletotrichum graminicola , Cercospora zeina , Fusarium verticillioides , and Fusarium virguliforme ) while limited expression was noted against abiotic stresses. Concisely, our results provide a platform for additional functional exploration of the ZmGLP genes against various environmental stresses.

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Section: Introductionmentioning

confidence: 99%

Molecular Characterization of Germin-like Protein Genes in Zea mays (ZmGLPs) Using Various In Silico Approaches

Ilyas¹,

Ali

Binjawhar

et al. 2023

ACS Omega

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“…No loss of activity was observed when the enzyme was incubated for 4 hours in the presence of a large excess of trypsin [ 21 ]. There is agreement that the monomer ranges from 23-26kDa depending on the nature of glycans added during post-translational modification; however, the structure of the active oligomer has been reported as a dimer [ 17 ] and tetramer [ 21 ], was also assumed to be a pentamer based on characterization of the wheat homologue [ 22 , 23 ], and most recently it has been identified as a hexamer based on X-ray crystallography studies [ 24 ]. EPR spectroscopy identified that the active form is a manganese-containing enzyme [ 19 , 25 ], and the formation of disulfide bonds are also essential for enzymatic activity [ 13 ].…”

Section: Introductionmentioning

confidence: 99%

“…EPR spectroscopy identified that the active form is a manganese-containing enzyme [ 19 , 25 ], and the formation of disulfide bonds are also essential for enzymatic activity [ 13 ]. No studies have specifically characterized glycans for the barley OxOx enzyme, but studies for the highly similar homologue in wheat indicate the presence of N-linked but not O-linked glycans [ 22 ].…”

Section: Introductionmentioning

confidence: 99%

Expression and secretion of glycosylated barley oxalate oxidase in Pichia pastoris

Donelan

Dominguez‐Gutierrez

et al. 2023

PLoS ONE

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Oxalate oxidase is an enzyme that degrades oxalate and is used in commercial urinary assays to measure oxalate levels. The objective of this study was to establish an enhanced expression system for secretion and purification of oxalate oxidase using Pichia pastoris. A codon optimized synthetic oxalate oxidase gene derived from Hordeum vulgare (barley) was generated and cloned into the pPICZα expression vector downstream of the N-terminal alpha factor secretion signal peptide sequence and used for expression in P. pastoris X-33 strain. A novel chimeric signal peptide consisting of the pre-OST1 sequence fused to pro-αpp8 containing several amino acid substitutions was also generated to enhance secretion. Active enzyme was purified to greater than 90% purity using Q-Sepharose anion exchange chromatography. The purified oxalate oxidase enzyme had an estimated Km value of 256μM, and activity was determined to be 10U/mg. We have developed an enhanced oxalate oxidase expression system and method for purification.

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“…Germins and GLPs share a well conserved β-barrel fold [1] and are typically glycoproteins. Differences in the extent of glycosylation are responsible for their presence as different isoforms, with variably modified forms associated with different subcellular localization [6,7] and tissue distribution [8]. Oryza sativa root germin like protein 1 (OsRGLP1) contains a putative N-glycosylation site (N-x-S/T; x¹P) [9], but the exact role of glycosylation in OsRGLP1 protein stability, folding, or activity is unknown.…”

Section: Introductionmentioning

confidence: 99%

N-glycosylation on Oryza sativa root germin-like protein 1 is conserved but not required for stability or activity

Rubbab

Pegg

Phung

et al. 2021

Biochemical and Biophysical Research Communications

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Covalently bonded and adventitious glycans in germin.

Cited by 69 publications

References 57 publications

Molecular Characterization of Germin-like Protein Genes in Zea mays (ZmGLPs) Using Various In Silico Approaches

Molecular Characterization of Germin-like Protein Genes in Zea mays (ZmGLPs) Using Various In Silico Approaches

Expression and secretion of glycosylated barley oxalate oxidase in Pichia pastoris

N-glycosylation on Oryza sativa root germin-like protein 1 is conserved but not required for stability or activity

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