The Detection and Quantitation of Free Desmosine and Isodesmosine in Human Urine and Their Peptide-Bound Forms in Sputum
Desmosine (D) and isodesmosine (I), the intramolecular crosslinking amino acids that occur in chains of elastin, have now been found in free form in human urine. Until now, these amino acids (Mr= 526) were found to occur in urine only as higher molecular weight (Mr= 1,000-1,500) peptides. Thus, the...
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| Published in | Proceedings of the National Academy of Sciences - PNAS Vol. 100; no. 22; pp. 12941 - 12943 |
|---|---|
| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
National Academy of Sciences
28.10.2003
National Acad Sciences |
| Subjects | |
| Online Access | Get full text |
| ISSN | 0027-8424 1091-6490 1091-6490 |
| DOI | 10.1073/pnas.2235344100 |
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| Abstract | Desmosine (D) and isodesmosine (I), the intramolecular crosslinking amino acids that occur in chains of elastin, have now been found in free form in human urine. Until now, these amino acids (Mr= 526) were found to occur in urine only as higher molecular weight (Mr= 1,000-1,500) peptides. Thus, the previously used analytical methods required, as the first step, acid hydrolysis of the urine at elevated temperature to liberate D and I from their peptides. The analytical method described here uses HPLC followed by electrospray ionization MS for the detection and quantitation of free D and I in unhydrolyzed urine. Identities of both D and I were established by their retention times on LC and by their mass ion at 526 atomic mass units, characteristic of each compound. The sensitivity of the method is 0.10 ng. The average values of free D and I in the urine of seven healthy subjects were 1.42 ± 1.16 and$1.39 \pm 1.04\>\mu g/g$of creatinine, respectively. After acid hydrolysis of the urine, the amounts of D and I were 8.67 ± 3.75 and$6.28 \pm 2.87\>\mu g/g$of creatinine, respectively. The method was also successfully used to measure peptide-bound D and I levels in the sputum of patients with chronic obstructive pulmonary disease. |
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| AbstractList | Desmosine (D) and isodesmosine (I), the intramolecular crosslinking amino acids that occur in chains of elastin, have now been found in free form in human urine. Until now, these amino acids (M(r) = 526) were found to occur in urine only as higher molecular weight (M (r) = 1,000-1,500) peptides. Thus, the previously used analytical methods required, as the first step, acid hydrolysis of the urine at elevated temperature to liberate D and I from their peptides. The analytical method described here uses HPLC followed by electrospray ionization MS for the detection and quantitation of free D and I in unhydrolyzed urine. Identities of both D and I were established by their retention times on LC and by their mass ion at 526 atomic mass units, characteristic of each compound. The sensitivity of the method is 0.10 ng. The average values of free D and I in the urine of seven healthy subjects were 1.42 +/- 1.16 and 1.39 +/- 1.04 microg/g of creatinine, respectively. After acid hydrolysis of the urine, the amounts of D and I were 8.67 +/- 3.75 and 6.28+/-2.87 microg/g of creatinine, respectively. The method was also successfully used to measure peptide-bound D and I levels in the sputum of patients with chronic obstructive pulmonary disease.Desmosine (D) and isodesmosine (I), the intramolecular crosslinking amino acids that occur in chains of elastin, have now been found in free form in human urine. Until now, these amino acids (M(r) = 526) were found to occur in urine only as higher molecular weight (M (r) = 1,000-1,500) peptides. Thus, the previously used analytical methods required, as the first step, acid hydrolysis of the urine at elevated temperature to liberate D and I from their peptides. The analytical method described here uses HPLC followed by electrospray ionization MS for the detection and quantitation of free D and I in unhydrolyzed urine. Identities of both D and I were established by their retention times on LC and by their mass ion at 526 atomic mass units, characteristic of each compound. The sensitivity of the method is 0.10 ng. The average values of free D and I in the urine of seven healthy subjects were 1.42 +/- 1.16 and 1.39 +/- 1.04 microg/g of creatinine, respectively. After acid hydrolysis of the urine, the amounts of D and I were 8.67 +/- 3.75 and 6.28+/-2.87 microg/g of creatinine, respectively. The method was also successfully used to measure peptide-bound D and I levels in the sputum of patients with chronic obstructive pulmonary disease. Desmosine (D) and isodesmosine (I), the intramolecular crosslinking amino acids that occur in chains of elastin, have now been found in free form in human urine. Until now, these amino acids ( M r = 526) were found to occur in urine only as higher molecular weight ( M r = 1,000–1,500) peptides. Thus, the previously used analytical methods required, as the first step, acid hydrolysis of the urine at elevated temperature to liberate D and I from their peptides. The analytical method described here uses HPLC followed by electrospray ionization MS for the detection and quantitation of free D and I in unhydrolyzed urine. Identities of both D and I were established by their retention times on LC and by their mass ion at 526 atomic mass units, characteristic of each compound. The sensitivity of the method is 0.10 ng. The average values of free D and I in the urine of seven healthy subjects were 1.42 ± 1.16 and 1.39 ± 1.04 μg/g of creatinine, respectively. After acid hydrolysis of the urine, the amounts of D and I were 8.67 ± 3.75 and 6.28 ± 2.87 μg/g of creatinine, respectively. The method was also successfully used to measure peptide-bound D and I levels in the sputum of patients with chronic obstructive pulmonary disease. Desmosine (D) and isodesmosine (I), the intramolecular crosslinking amino acids that occur in chains of elastin, have now been found in free form in human urine. Until now, these amino acids (Mr= 526) were found to occur in urine only as higher molecular weight (Mr= 1,000-1,500) peptides. Thus, the previously used analytical methods required, as the first step, acid hydrolysis of the urine at elevated temperature to liberate D and I from their peptides. The analytical method described here uses HPLC followed by electrospray ionization MS for the detection and quantitation of free D and I in unhydrolyzed urine. Identities of both D and I were established by their retention times on LC and by their mass ion at 526 atomic mass units, characteristic of each compound. The sensitivity of the method is 0.10 ng. The average values of free D and I in the urine of seven healthy subjects were 1.42 ± 1.16 and$1.39 \pm 1.04\>\mu g/g$of creatinine, respectively. After acid hydrolysis of the urine, the amounts of D and I were 8.67 ± 3.75 and$6.28 \pm 2.87\>\mu g/g$of creatinine, respectively. The method was also successfully used to measure peptide-bound D and I levels in the sputum of patients with chronic obstructive pulmonary disease. Desmosine (D) and isodesmosine (I), the intramolecular crosslinking amino acids that occur in chains of elastin, have now been found in free form in human urine. Until now, these amino acids (Mr = 526) were found to occur in urine only as higher molecular weight peptides. Thus, the previously used analytical methods required, as the first step, acid hydrolysis of the urine at elevated temperature to liberate D and I from their peptides. The analytical method described here uses HPLC followed by electrospray ionization MS for the detection and quantitation of free D and I in unhydrolyzed urine. Identities of both D and I were established by their retention times on LC and by their mass ion at 526 atomic mass units, characteristic of each compound. The sensitivity of the method is 0.10 ng. The average values of free D and I in the urine of seven healthy subjects were 1.42 plus or minus 1.16 and 1.39 plus or minus 1.04 gg of creatinine, respectively. After acid hydrolysis of the urine, the amounts of D and I were 8.67 plus or minus 2.87 gg of creatinine, respectively. The method was also successfully used to measure peptide-bound D and I levels in the sputum of patients with chronic obstructive pulmonary disease. [PUBLICATION ABSTRACT] Desmosine (D) and isodesmosine (I), the intramolecular crosslinking amino acids that occur in chains of elastin, have now been found in free form in human urine. Until now, these amino acids (M(r) = 526) were found to occur in urine only as higher molecular weight (M (r) = 1,000-1,500) peptides. Thus, the previously used analytical methods required, as the first step, acid hydrolysis of the urine at elevated temperature to liberate D and I from their peptides. The analytical method described here uses HPLC followed by electrospray ionization MS for the detection and quantitation of free D and I in unhydrolyzed urine. Identities of both D and I were established by their retention times on LC and by their mass ion at 526 atomic mass units, characteristic of each compound. The sensitivity of the method is 0.10 ng. The average values of free D and I in the urine of seven healthy subjects were 1.42 +/- 1.16 and 1.39 +/- 1.04 microg/g of creatinine, respectively. After acid hydrolysis of the urine, the amounts of D and I were 8.67 +/- 3.75 and 6.28+/-2.87 microg/g of creatinine, respectively. The method was also successfully used to measure peptide-bound D and I levels in the sputum of patients with chronic obstructive pulmonary disease. Desmosine (D) and isodesmosine (I), the intramolecular crosslinking amino acids that occur in chains of elastin, have now been found in free form in human urine. Until now, these amino acids ( M r = 526) were found to occur in urine only as higher molecular weight ( M r = 1,000–1,500) peptides. Thus, the previously used analytical methods required, as the first step, acid hydrolysis of the urine at elevated temperature to liberate D and I from their peptides. The analytical method described here uses HPLC followed by electrospray ionization MS for the detection and quantitation of free D and I in unhydrolyzed urine. Identities of both D and I were established by their retention times on LC and by their mass ion at 526 atomic mass units, characteristic of each compound. The sensitivity of the method is 0.10 ng. The average values of free D and I in the urine of seven healthy subjects were 1.42 ± 1.16 and 1.39 ± 1.04 μg/g of creatinine, respectively. After acid hydrolysis of the urine, the amounts of D and I were 8.67 ± 3.75 and 6.28 ± 2.87 μg/g of creatinine, respectively. The method was also successfully used to measure peptide-bound D and I levels in the sputum of patients with chronic obstructive pulmonary disease. Desmosine (D) and isodesmosine (I), the intramolecular crosslinking amino acids that occur in chains of elastin, have now been found in free form in human urine. Until now, these amino acids (Mr = 526) were found to occur in urine only as higher molecular weight (Mr = 1,000–1,500) peptides. Thus, the previously used analytical methods required, as the first step, acid hydrolysis of the urine at elevated temperature to liberate D and I from their peptides. The analytical method described here uses HPLC followed by electrospray ionization MS for the detection and quantitation of free D and I in unhydrolyzed urine. Identities of both D and I were established by their retention times on LC and by their mass ion at 526 atomic mass units, characteristic of each compound. The sensitivity of the method is 0.10 ng. The average values of free D and I in the urine of seven healthy subjects were 1.42 ± 1.16 and 1.39 ± 1.04 μg/g of creatinine, respectively. After acid hydrolysis of the urine, the amounts of D and I were 8.67 ± 3.75 and 6.28 ± 2.87 μg/g of creatinine, respectively. The method was also successfully used to measure peptide-bound D and I levels in the sputum of patients with chronic obstructive pulmonary disease. |
| Author | Ma, Shuren Lieberman, Seymour Turino, Gerard M. Lin, Yong Y. |
| AuthorAffiliation | James P. Mara Center for Lung Disease, Department of Medicine, St. Luke's/Roosevelt Hospital Center, Columbia University College of Physicians and Surgeons, New York, NY 10019 |
| AuthorAffiliation_xml | – name: James P. Mara Center for Lung Disease, Department of Medicine, St. Luke's/Roosevelt Hospital Center, Columbia University College of Physicians and Surgeons, New York, NY 10019 |
| Author_xml | – sequence: 1 givenname: Shuren surname: Ma fullname: Ma, Shuren – sequence: 2 givenname: Seymour surname: Lieberman fullname: Lieberman, Seymour – sequence: 3 givenname: Gerard M. surname: Turino fullname: Turino, Gerard M. – sequence: 4 givenname: Yong Y. surname: Lin fullname: Lin, Yong Y. |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/14563926$$D View this record in MEDLINE/PubMed |
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| Notes | SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 14 ObjectType-Article-1 ObjectType-Feature-2 content type line 23 Contributed by Seymour Lieberman, August 20, 2003 Abbreviations: D, desmosine; I, isodesmosine; ESIMS, electrospray ionization MS; SIR, selected ion recording; COPD, chronic obstructive pulmonary disease. |
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| Snippet | Desmosine (D) and isodesmosine (I), the intramolecular crosslinking amino acids that occur in chains of elastin, have now been found in free form in human... |
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| SubjectTerms | Acid hydrolysis Adult Aged Amino acids Analytical methods Biological Sciences Biomarkers - analysis Biomarkers - urine Chronic obstructive pulmonary disease Creatinine - analysis Creatinine - urine Desmosine - analogs & derivatives Desmosine - analysis Desmosine - urine Electric potential Female High temperature Humans Hydrolysis Ionization Ions Isodesmosine - analogs & derivatives Isodesmosine - analysis Isodesmosine - urine Isomers Liquid chromatography Lung diseases Male Mass Spectrometry Medical research Middle Aged Peptides Pulmonary Disease, Chronic Obstructive - metabolism Pulmonary Disease, Chronic Obstructive - urine Sputum Sputum - chemistry Urine |
| Title | The Detection and Quantitation of Free Desmosine and Isodesmosine in Human Urine and Their Peptide-Bound Forms in Sputum |
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