Amino acid sequence of a tryptic peptide of human thyroglobulin reactive with sera of patients with thyroid diseases

Ali M. Saboori, Noel R. Rose, C. Lynne Burek

Research output: Contribution to journalArticle

Abstract

Autoantibodies to human thyroglobulin (hTg) are found in the sera of many patients with thyroid diseases. To localize epitopes recognized by these autoantibodies, hTg was incubated with tryspin for 4 hours at 37°C under non-reducing conditions, releasing peptides from hTg in their natural conformation. These peptides were then analyzed by western immunoblot using either autoantibodies from patients with autoimmune thyroiditis or murine monoclonal antibodies (mAb) produced against hTg. The autoantibodies reacted primarily with two low molecular weight peptides with apparent molecular weights (MWap) of 15 and 20 kDa. The pattern of tryptic peptides recognized by these autoantibodies resembled that of one of the mAbs (137C1), as shown by immunoblots in either one or two dimensional SDS-PAGE. To characterize these peptides further, they were separated by a high performance liquid chromatography (HPLC). The column separated the 4-hour tryptic digest of hTg into multiple peptide peaks. Further analysis by SDS-PAGE showed that one of these peaks contained the 15 kDa peptide. The 15 amino acid sequence at the amino-terminus of this peptide was determined. This amino acid sequence (KVPTFATPWPDFVPR) corresponds to a unique sequence near the carboxyl-terminal end of hTg, starting with amino acid 2657, The size of the peptide indicates that it extends to the carboxyl-terminal end of hTg. This fragment contains one of the antigenic sites of hTg that binds autoantibodies from patients with autoimmune thyroid disease.

Original languageEnglish (US)
Pages (from-to)87-94
Number of pages8
JournalAutoimmunity
Volume22
Issue number2
DOIs
StatePublished - 1995

Fingerprint

Thyroglobulin
Thyroid Diseases
Amino Acid Sequence
Autoantibodies
Peptides
Serum
Polyacrylamide Gel Electrophoresis
Molecular Weight
Autoimmune Thyroiditis
Autoimmune Diseases
Epitopes
Western Blotting
High Pressure Liquid Chromatography
Monoclonal Antibodies
Amino Acids

Keywords

  • Autoantibodies
  • Thyroglobulin
  • Thyroiditis

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Amino acid sequence of a tryptic peptide of human thyroglobulin reactive with sera of patients with thyroid diseases. / Saboori, Ali M.; Rose, Noel R.; Burek, C. Lynne.

In: Autoimmunity, Vol. 22, No. 2, 1995, p. 87-94.

Research output: Contribution to journalArticle

Saboori, Ali M. ; Rose, Noel R. ; Burek, C. Lynne. / Amino acid sequence of a tryptic peptide of human thyroglobulin reactive with sera of patients with thyroid diseases. In: Autoimmunity. 1995 ; Vol. 22, No. 2. pp. 87-94.
@article{78d1eef203ed4d75bc0e33e2f7fb5dfd,
title = "Amino acid sequence of a tryptic peptide of human thyroglobulin reactive with sera of patients with thyroid diseases",
abstract = "Autoantibodies to human thyroglobulin (hTg) are found in the sera of many patients with thyroid diseases. To localize epitopes recognized by these autoantibodies, hTg was incubated with tryspin for 4 hours at 37°C under non-reducing conditions, releasing peptides from hTg in their natural conformation. These peptides were then analyzed by western immunoblot using either autoantibodies from patients with autoimmune thyroiditis or murine monoclonal antibodies (mAb) produced against hTg. The autoantibodies reacted primarily with two low molecular weight peptides with apparent molecular weights (MWap) of 15 and 20 kDa. The pattern of tryptic peptides recognized by these autoantibodies resembled that of one of the mAbs (137C1), as shown by immunoblots in either one or two dimensional SDS-PAGE. To characterize these peptides further, they were separated by a high performance liquid chromatography (HPLC). The column separated the 4-hour tryptic digest of hTg into multiple peptide peaks. Further analysis by SDS-PAGE showed that one of these peaks contained the 15 kDa peptide. The 15 amino acid sequence at the amino-terminus of this peptide was determined. This amino acid sequence (KVPTFATPWPDFVPR) corresponds to a unique sequence near the carboxyl-terminal end of hTg, starting with amino acid 2657, The size of the peptide indicates that it extends to the carboxyl-terminal end of hTg. This fragment contains one of the antigenic sites of hTg that binds autoantibodies from patients with autoimmune thyroid disease.",
keywords = "Autoantibodies, Thyroglobulin, Thyroiditis",
author = "Saboori, {Ali M.} and Rose, {Noel R.} and Burek, {C. Lynne}",
year = "1995",
doi = "10.3109/08916939508995304",
language = "English (US)",
volume = "22",
pages = "87--94",
journal = "Autoimmunity",
issn = "0891-6934",
publisher = "Informa Healthcare",
number = "2",

}

TY - JOUR

T1 - Amino acid sequence of a tryptic peptide of human thyroglobulin reactive with sera of patients with thyroid diseases

AU - Saboori, Ali M.

AU - Rose, Noel R.

AU - Burek, C. Lynne

PY - 1995

Y1 - 1995

N2 - Autoantibodies to human thyroglobulin (hTg) are found in the sera of many patients with thyroid diseases. To localize epitopes recognized by these autoantibodies, hTg was incubated with tryspin for 4 hours at 37°C under non-reducing conditions, releasing peptides from hTg in their natural conformation. These peptides were then analyzed by western immunoblot using either autoantibodies from patients with autoimmune thyroiditis or murine monoclonal antibodies (mAb) produced against hTg. The autoantibodies reacted primarily with two low molecular weight peptides with apparent molecular weights (MWap) of 15 and 20 kDa. The pattern of tryptic peptides recognized by these autoantibodies resembled that of one of the mAbs (137C1), as shown by immunoblots in either one or two dimensional SDS-PAGE. To characterize these peptides further, they were separated by a high performance liquid chromatography (HPLC). The column separated the 4-hour tryptic digest of hTg into multiple peptide peaks. Further analysis by SDS-PAGE showed that one of these peaks contained the 15 kDa peptide. The 15 amino acid sequence at the amino-terminus of this peptide was determined. This amino acid sequence (KVPTFATPWPDFVPR) corresponds to a unique sequence near the carboxyl-terminal end of hTg, starting with amino acid 2657, The size of the peptide indicates that it extends to the carboxyl-terminal end of hTg. This fragment contains one of the antigenic sites of hTg that binds autoantibodies from patients with autoimmune thyroid disease.

AB - Autoantibodies to human thyroglobulin (hTg) are found in the sera of many patients with thyroid diseases. To localize epitopes recognized by these autoantibodies, hTg was incubated with tryspin for 4 hours at 37°C under non-reducing conditions, releasing peptides from hTg in their natural conformation. These peptides were then analyzed by western immunoblot using either autoantibodies from patients with autoimmune thyroiditis or murine monoclonal antibodies (mAb) produced against hTg. The autoantibodies reacted primarily with two low molecular weight peptides with apparent molecular weights (MWap) of 15 and 20 kDa. The pattern of tryptic peptides recognized by these autoantibodies resembled that of one of the mAbs (137C1), as shown by immunoblots in either one or two dimensional SDS-PAGE. To characterize these peptides further, they were separated by a high performance liquid chromatography (HPLC). The column separated the 4-hour tryptic digest of hTg into multiple peptide peaks. Further analysis by SDS-PAGE showed that one of these peaks contained the 15 kDa peptide. The 15 amino acid sequence at the amino-terminus of this peptide was determined. This amino acid sequence (KVPTFATPWPDFVPR) corresponds to a unique sequence near the carboxyl-terminal end of hTg, starting with amino acid 2657, The size of the peptide indicates that it extends to the carboxyl-terminal end of hTg. This fragment contains one of the antigenic sites of hTg that binds autoantibodies from patients with autoimmune thyroid disease.

KW - Autoantibodies

KW - Thyroglobulin

KW - Thyroiditis

UR - http://www.scopus.com/inward/record.url?scp=0029552180&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029552180&partnerID=8YFLogxK

U2 - 10.3109/08916939508995304

DO - 10.3109/08916939508995304

M3 - Article

C2 - 8722578

AN - SCOPUS:0029552180

VL - 22

SP - 87

EP - 94

JO - Autoimmunity

JF - Autoimmunity

SN - 0891-6934

IS - 2

ER -