Identification of the novel autoantigen candidate Rab GDP dissociation inhibitor alpha in isolated adrenocorticotropin deficiency

Atsushi Kiyota, Shintaro Iwama, Yoshihisa Sugimura, Seiji Takeuchi, Hiroshi Takagi, Naoko Iwata, Kohtaro Nakashima, Haruyuki Suzuki, Tomoki Nishioka, Takuya Kato, Atsushi Enomoto, Hiroshi Arima, Kozo Kaibuchi, Yutaka Oiso

Research output: Contribution to journalArticle

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Abstract

Isolated adrenocorticotropin deficiency (IAD) is characterized by low or absent adrenocorticotropic hormone (ACTH) production. IAD is presumed to be caused in part by an autoimmune mechanism, and several lines of evidence have suggested the presence of anti-pituitary antibodies in IAD. However, the exact autoantigens remain unknown. The present study was designed to identify the autoantigen(s) in IAD using chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Rat anterior pituitary lysate was subjected to SDS-PAGE, and immunoblotting was performed using the sera from two patients with IAD and from a healthy subject. The bands detected by the patient serum samples, but not by the healthy subject sample, were excised, in-gel digested using trypsin, and subjected to LC-MS/MS analysis. On immunoblots, a 51- kDa band in the insoluble pellet was detected by the sera from the IAD patients but not from the healthy subject. Mass spectrometric analysis revealed the 51-kDa band contained Rab guanine nucleotide dissociation inhibitor (GDI) alpha. Consistent with the mass spectrometric analysis, a recombinant full-length human Rab GDI alpha was recognized by the two IAD patient samples but not by the healthy subject sample using immunoblotting. In total, anti-Rab GDI alpha antibodies were detected in serum samples from three of five patients with IAD (60%) but were absent in 5 healthy subjects. In addition, Rab GDI alpha was expressed in the anterior pituitary. In conclusion, it appears that Rab GDI alpha is a candidate autoantigen involved in IAD, and that anti-Rab GDI alpha antibodies are present predominantly in patients with IAD.

Original languageEnglish
Pages (from-to)153-160
Number of pages8
JournalEndocrine Journal
Volume62
Issue number2
DOIs
Publication statusPublished - 01-01-2015

Fingerprint

Guanine Nucleotide Dissociation Inhibitors
Autoantigens
Adrenocorticotropic Hormone
Healthy Volunteers
Serum
Immunoblotting
Antibodies
Tandem Mass Spectrometry
Trypsin
GDP dissociation inhibitor 1
Chromatography
Polyacrylamide Gel Electrophoresis
Anti-Idiotypic Antibodies

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology

Cite this

Kiyota, Atsushi ; Iwama, Shintaro ; Sugimura, Yoshihisa ; Takeuchi, Seiji ; Takagi, Hiroshi ; Iwata, Naoko ; Nakashima, Kohtaro ; Suzuki, Haruyuki ; Nishioka, Tomoki ; Kato, Takuya ; Enomoto, Atsushi ; Arima, Hiroshi ; Kaibuchi, Kozo ; Oiso, Yutaka. / Identification of the novel autoantigen candidate Rab GDP dissociation inhibitor alpha in isolated adrenocorticotropin deficiency. In: Endocrine Journal. 2015 ; Vol. 62, No. 2. pp. 153-160.
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abstract = "Isolated adrenocorticotropin deficiency (IAD) is characterized by low or absent adrenocorticotropic hormone (ACTH) production. IAD is presumed to be caused in part by an autoimmune mechanism, and several lines of evidence have suggested the presence of anti-pituitary antibodies in IAD. However, the exact autoantigens remain unknown. The present study was designed to identify the autoantigen(s) in IAD using chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Rat anterior pituitary lysate was subjected to SDS-PAGE, and immunoblotting was performed using the sera from two patients with IAD and from a healthy subject. The bands detected by the patient serum samples, but not by the healthy subject sample, were excised, in-gel digested using trypsin, and subjected to LC-MS/MS analysis. On immunoblots, a 51- kDa band in the insoluble pellet was detected by the sera from the IAD patients but not from the healthy subject. Mass spectrometric analysis revealed the 51-kDa band contained Rab guanine nucleotide dissociation inhibitor (GDI) alpha. Consistent with the mass spectrometric analysis, a recombinant full-length human Rab GDI alpha was recognized by the two IAD patient samples but not by the healthy subject sample using immunoblotting. In total, anti-Rab GDI alpha antibodies were detected in serum samples from three of five patients with IAD (60{\%}) but were absent in 5 healthy subjects. In addition, Rab GDI alpha was expressed in the anterior pituitary. In conclusion, it appears that Rab GDI alpha is a candidate autoantigen involved in IAD, and that anti-Rab GDI alpha antibodies are present predominantly in patients with IAD.",
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Kiyota, A, Iwama, S, Sugimura, Y, Takeuchi, S, Takagi, H, Iwata, N, Nakashima, K, Suzuki, H, Nishioka, T, Kato, T, Enomoto, A, Arima, H, Kaibuchi, K & Oiso, Y 2015, 'Identification of the novel autoantigen candidate Rab GDP dissociation inhibitor alpha in isolated adrenocorticotropin deficiency', Endocrine Journal, vol. 62, no. 2, pp. 153-160. https://doi.org/10.1507/endocrj.EJ14-0369

Identification of the novel autoantigen candidate Rab GDP dissociation inhibitor alpha in isolated adrenocorticotropin deficiency. / Kiyota, Atsushi; Iwama, Shintaro; Sugimura, Yoshihisa; Takeuchi, Seiji; Takagi, Hiroshi; Iwata, Naoko; Nakashima, Kohtaro; Suzuki, Haruyuki; Nishioka, Tomoki; Kato, Takuya; Enomoto, Atsushi; Arima, Hiroshi; Kaibuchi, Kozo; Oiso, Yutaka.

In: Endocrine Journal, Vol. 62, No. 2, 01.01.2015, p. 153-160.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification of the novel autoantigen candidate Rab GDP dissociation inhibitor alpha in isolated adrenocorticotropin deficiency

AU - Kiyota, Atsushi

AU - Iwama, Shintaro

AU - Sugimura, Yoshihisa

AU - Takeuchi, Seiji

AU - Takagi, Hiroshi

AU - Iwata, Naoko

AU - Nakashima, Kohtaro

AU - Suzuki, Haruyuki

AU - Nishioka, Tomoki

AU - Kato, Takuya

AU - Enomoto, Atsushi

AU - Arima, Hiroshi

AU - Kaibuchi, Kozo

AU - Oiso, Yutaka

PY - 2015/1/1

Y1 - 2015/1/1

N2 - Isolated adrenocorticotropin deficiency (IAD) is characterized by low or absent adrenocorticotropic hormone (ACTH) production. IAD is presumed to be caused in part by an autoimmune mechanism, and several lines of evidence have suggested the presence of anti-pituitary antibodies in IAD. However, the exact autoantigens remain unknown. The present study was designed to identify the autoantigen(s) in IAD using chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Rat anterior pituitary lysate was subjected to SDS-PAGE, and immunoblotting was performed using the sera from two patients with IAD and from a healthy subject. The bands detected by the patient serum samples, but not by the healthy subject sample, were excised, in-gel digested using trypsin, and subjected to LC-MS/MS analysis. On immunoblots, a 51- kDa band in the insoluble pellet was detected by the sera from the IAD patients but not from the healthy subject. Mass spectrometric analysis revealed the 51-kDa band contained Rab guanine nucleotide dissociation inhibitor (GDI) alpha. Consistent with the mass spectrometric analysis, a recombinant full-length human Rab GDI alpha was recognized by the two IAD patient samples but not by the healthy subject sample using immunoblotting. In total, anti-Rab GDI alpha antibodies were detected in serum samples from three of five patients with IAD (60%) but were absent in 5 healthy subjects. In addition, Rab GDI alpha was expressed in the anterior pituitary. In conclusion, it appears that Rab GDI alpha is a candidate autoantigen involved in IAD, and that anti-Rab GDI alpha antibodies are present predominantly in patients with IAD.

AB - Isolated adrenocorticotropin deficiency (IAD) is characterized by low or absent adrenocorticotropic hormone (ACTH) production. IAD is presumed to be caused in part by an autoimmune mechanism, and several lines of evidence have suggested the presence of anti-pituitary antibodies in IAD. However, the exact autoantigens remain unknown. The present study was designed to identify the autoantigen(s) in IAD using chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Rat anterior pituitary lysate was subjected to SDS-PAGE, and immunoblotting was performed using the sera from two patients with IAD and from a healthy subject. The bands detected by the patient serum samples, but not by the healthy subject sample, were excised, in-gel digested using trypsin, and subjected to LC-MS/MS analysis. On immunoblots, a 51- kDa band in the insoluble pellet was detected by the sera from the IAD patients but not from the healthy subject. Mass spectrometric analysis revealed the 51-kDa band contained Rab guanine nucleotide dissociation inhibitor (GDI) alpha. Consistent with the mass spectrometric analysis, a recombinant full-length human Rab GDI alpha was recognized by the two IAD patient samples but not by the healthy subject sample using immunoblotting. In total, anti-Rab GDI alpha antibodies were detected in serum samples from three of five patients with IAD (60%) but were absent in 5 healthy subjects. In addition, Rab GDI alpha was expressed in the anterior pituitary. In conclusion, it appears that Rab GDI alpha is a candidate autoantigen involved in IAD, and that anti-Rab GDI alpha antibodies are present predominantly in patients with IAD.

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