Distribution of storage proteins in low-glutelin rice seed determined using a fluorescent antibody

Furukawa, Sachiko; Mizuma, Tomochika; Kiyokawa, Yoshifumi; Masumura, Takehiro; Tanaka, Kunisuke; Wakai, Yoshinori

doi:10.1016/s1389-1723(03)70133-9

Cited by 37 publications

(35 citation statements)

References 14 publications

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“…After blocking, the membranes were incubated for 1 h with antibodies against 23G (diluted with the blocking solution 1 : 20,000, v : v) or 13P (diluted 1 : 20,000, v : v) at room temperature (11). It was checked in the previous report that these antibodies used for WB analysis did not detect endogenous proteins in GI contents of rats (6).…”

Section: Inmentioning

confidence: 99%

In Vivo Digestibility of Rice Prolamin/Protein Body-I Particle Is Decreased by Cooking

Kubota

Saito

Masumura

et al. 2014

J Nutr Sci Vitaminol

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Rice is a staple cereal and a major source of both protein and energy in most Asian countries. Thus, the true digestibility of cooked rice protein is very important for human nutrition. The proteins in rice endosperm consist mainly of storage proteins, e.g., glutelin, prolamin and globulin. Two major proteins, glutelin and prolamin, can be separated into several protein bands by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Glutelin separates into three bands: proglutelin (57 kDa), an acidic subunit (37-39 kDa), and a basic subunit (22-23 kDa) (1). Prolamin also separates into three bands: 16, 13 and 10 kDa prolamin (2). These storage proteins accumulate in organelles called protein bodies (PBs). Rice endosperm has two different types of PBs; prolamin accumulates in type-I protein bodies (PB-I), while glutelin and globulin accumulate in type-II protein bodies (PB-II) (3). Tanaka et al. (4) reported that fecal protein particles (FPP), such as PBs, were excreted in the feces of humans who had ingested cooked rice (CR). Ogawa et al. (2) demonstrated that the 13 kDa prolamin band in raw rice (RR) remained after pepsin digestion, although the glutelin subunit bands (37-39 and 22-23 kDa) disappeared. These results imply that prolamin is an indigestible protein. However, we recently reported that structural changes induced by alkali extraction can improve the in vivo digestibility of prolamin (5, 6). A study by Eggum et al. showed that cooking decreased the true protein digestibility (TD) of rice from 99.7 to 88.6% (7). However, no reports have directly demonstrated whether rice prolamin is rendered indigestible in vivo by cooking.In the present study, we monitored the gastrointestinal (GI) transit and fecal excretion of rice glutelin and prolamin in rats by using anti-23 kDa glutelin (23G) and anti-13 kDa prolamin (13P) antibodies to estimate their digestibility in CR and RR. Additionally, morphological observations of PBs in the feces of rats fed the CR or RR diets were conducted using electron microscopy to confirm the effects of cooking on PB structure. Although the digestibility of 23G was not affected by cooking, the digestibility of 13P was clearly reduced. Almost intact PB-Is were observed in the feces of rats fed the CR diet. Our results clearly indicate that rice prolamin is not indigestible in its raw form, but is rendered indigestible by cooking. Materials and MethodsMaterials and animal experiment. Rice (Koshihikari, 10% polished fraction) was purchased from Niigata Seifun Co., Ltd. (Tainai, Japan). RR flour was prepared using a jet mill (KV-3-4S, Yakushinkikai Seisakusho, Japan). CR flour was prepared by rinsing the rice first and then cooking it in twice its volume of water in an electric rice cooker (JAG-A100 TM, Tiger Co., Ltd., Osaka, Japan). Summary Rice has storage proteins, e.g., glutelin, globulin and prolamin, in the seeds, which are used as nitrogen sources during germination. Rice prolamin has been reported to be an indigestible protein that decreases the nutritional ...

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Section: Inmentioning

confidence: 99%

In Vivo Digestibility of Rice Prolamin/Protein Body-I Particle Is Decreased by Cooking

Kubota

Saito

Masumura

et al. 2014

J Nutr Sci Vitaminol

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“…5, Ohdaira et al, 2009). Even in a comprehensive study by Furukawa et al (2003), the observation site examined with fl uorescence microscopy was a part of the outer region of a 30% milled rice grain.…”

Section: Discussionmentioning

confidence: 99%

“…Furukawa et al (2003) observed rice sections of low-glutelin cultivars using fluorescence-labeled antibodies of 13 kDa prolamin and 23 kDa glutelin, which are, respectively, major components of protein body type I and protein body type II. They reported that a large amount of 13 kDa prolamin existed in the outer region of rice grains after 30% milling in lowglutelin cultivars, in addition to the outer regions of brown rice.…”

mentioning

confidence: 99%

Analysis of Storage Protein Distribution in Rice Grain of Seed-Protein Mutant Cultivars by Immunofluorescence Microscopy

Ohdaira

Masumura

Nakatsuka

et al. 2011

Plant Production Science

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“…Then total proteins of this section were stained with Coomassie brilliant blue (CBB) for 5 min. For immunocytochemistry, anti-13 kDa prolamin antibodies 5) were conjugated with fluorescent dye Cy3 using a FluoroLink-Ab Cy3 labeling kit (GE Healthcare, Buckinghamshire, UK). The fixed sections were treated with blocking solution of 1% bovine serum albumin in 0.1 M phosphate buffer (pH 7.2) for 30 min at room temperature.…”

Section: Thin Frozen Film Methods For Visualization Of Storage Proteinmentioning

confidence: 99%

Thin Frozen Film Method for Visualization of Storage Proteins in Mature Rice Grains

Saito¹,

Nakatsuka²,

Shigemitsu³

et al. 2008

Bioscience, Biotechnology, and Biochemistry

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Distribution of storage proteins in low-glutelin rice seed determined using a fluorescent antibody

Cited by 37 publications

References 14 publications

In Vivo Digestibility of Rice Prolamin/Protein Body-I Particle Is Decreased by Cooking

In Vivo Digestibility of Rice Prolamin/Protein Body-I Particle Is Decreased by Cooking

Analysis of Storage Protein Distribution in Rice Grain of Seed-Protein Mutant Cultivars by Immunofluorescence Microscopy

Thin Frozen Film Method for Visualization of Storage Proteins in Mature Rice Grains

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