Dataset Information

Catalytic and Structural Characterization of a Browning-Related Protein in Oriental Sweet Melon (Cucumis Melo var. Makuwa Makino).

ABSTRACT: Polyphenol oxidase (PPO) in plants plays an important role in browning reactions and may affect the quality of sweet melon products. In this study, a browning-related protein (BRP) with PPO activity was partially purified from oriental sweet melon (Cucumis melo var. makuwa Makino) by salt precipitation and column chromatography. The BRP possessed a high degree of identity with several chitinase proteins, particularly defense-related proteins, by MS identification. Pyrogallol was determined as the most appropriate substrate for BRP (Km = 0.04278 M). BRP exhibited extreme resistance under alkaline and high temperature conditions when pyrogallol was used as substrate. Polyacrylamide gel electrophoresis (PAGE) analysis indicated that BRP was a homo-dimer of two subunits and had a molecular weight of 37 kDa. Structural analysis indicated that the ?-helix was the dominant conformation of BRP. The active site of the protein might be buried deeply in the protein, and BRP might be monodispersed in an aqueous system.

SUBMITTER: Liu S

PROVIDER: S-EPMC6116579 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Publications

Catalytic and Structural Characterization of a Browning-Related Protein in Oriental Sweet Melon (Cucumis Melo var. Makuwa Makino).

Liu Siyu S Murtaza Ayesha A Liu Yan Y Hu Wanfeng W Xu Xiaoyun X Pan Siyi S

Frontiers in chemistry 20180823

Polyphenol oxidase (PPO) in plants plays an important role in browning reactions and may affect the quality of sweet melon products. In this study, a browning-related protein (BRP) with PPO activity was partially purified from oriental sweet melon (Cucumis melo var. makuwa Makino) by salt precipitation and column chromatography. The BRP possessed a high degree of identity with several chitinase proteins, particularly defense-related proteins, by MS identification. Pyrogallol was de ...[more]

PMID: 30191148

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Project description:BACKGROUND:Lipoxygenases (LOXs) play significant roles in abiotic stress responses, and identification of LOX gene promoter function can make an important contribution to elucidating resistance mechanisms. Here, we cloned the CmLOX08 promoter of melon (Cucumis melo) and identified the main promoter regions regulating transcription in response to signalling molecules and abiotic stresses. RESULTS:The 2054-bp promoter region of CmLOX08 from melon leaves was cloned, and bioinformatic analysis revealed that it harbours numerous cis-regulatory elements associated with signalling molecules and abiotic stress. Five 5'-deletion fragments obtained from the CmLOX08 promoter-2054 (LP1), 1639 (LP2), 1284 (LP3), 1047 (LP4), and 418?bp (LP5)-were fused with a GUS reporter gene and used for tobacco transient assays. Deletion analysis revealed that in response to abscisic acid, salicylic acid, and hydrogen peroxide, the GUS activity of LP1 was significantly higher than that of the mock-treated control and LP2, indicating that the -?2054- to -?1639-bp region positively regulates expression induced by these signalling molecules. However, no deletion fragment GUS activity was induced by methyl jasmonate. In response to salt, drought, and wounding treatments, LP1, LP2, and LP4 promoted significantly higher GUS expression compared with the control. Among all deletion fragments, LP4 showed the highest GUS expression, indicating that -?1047 to -?1?bp is the major region regulating promoter activity and that the -?1047 to -?418-bp region positively regulates expression induced by salt, drought, and wounding, whereas the -?1284 to -?1047-bp region is a negative regulatory segment. Interestingly, although the GUS activity of LP1 and LP2 was not affected by temperature changes, that of LP3 was significantly induced by heat, indicating that the -?1284- to -?1-bp region is a core sequence responding to heat and the -?2054- to -?1284-bp region negatively regulates expression induced by heat. Similarly, the -?1047- to -?1-bp region is the main sequence responding to cold, whereas the -?2054- to -?1047-bp region negatively regulates expression induced by cold. CONCLUSIONS:We cloned the CmLOX08 promoter and demonstrated that it is a signalling molecule/stress-inducible promoter. Furthermore, we identified core and positive/negative regulatory regions responding to three signalling molecules and five abiotic stresses.

Dataset Information

Catalytic and Structural Characterization of a Browning-Related Protein in Oriental Sweet Melon (Cucumis Melo var. Makuwa Makino).

Publications

Catalytic and Structural Characterization of a Browning-Related Protein in Oriental Sweet Melon (<i>Cucumis Melo</i> var. <i>Makuwa</i> Makino).

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