Synthesis and biological activity of a biotinylated vasopressin analog

Ward H. Lutz; James M. Londowski; Rajiv Kumar

doi:10.1016/0196-9781(90)90181-4

Synthesis and biological activity of a biotinylated vasopressin analog

Ward H. Lutz, James M. Londowski, Rajiv Kumar

Nephrology and Hypertension

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

Abstract

To study vasopressin receptor-mediated endocytosis using electronmicroscopy methods and to develop avidin affinity columns for receptor purification, we synthesized and tested the biological properties of a biotinylated vasopressin (VP) analog [1-(2-mercapto) propionic acid] 8-[lysine-N⁶-biotin] VP (B-MLVP). B-MLVP was prepared by coupling biotin to the epsilon amine of the lysine residue in [1-(2-mercapto) propionic acid] 8-(lysine) VP (MLVP). The structure of HPLC purified B-MLVP was confirmed by fast atom bombardment mass spectrometry. B-MLVP effectively competed for arginine vasopressin (AVP) binding sites in canine renal plasma membranes on the surface of LLC-PK₁ kidney cells. Dissociation constants of 15 nM and 202 nM were calculated from the results of competition binding assays conducted with membranes and cells, respectively. B-MLVP stimulated adenylate cyclase activity and elevated cellular 3′,5′,cyclic-AMP (cAMP) content in a manner similar to AVP, indicating it is an agonist of VP action in renal tissue. These observations indicate that B-MLVP is an agonist of VP action and may be used to study renal VP receptors by employing avidin coupled to various reporter groups.

Original language	English (US)
Pages (from-to)	687-691
Number of pages	5
Journal	Peptides
Volume	11
Issue number	4
DOIs	https://doi.org/10.1016/0196-9781(90)90181-4
State	Published - 1990

Keywords

Biotin
Biotinylated analog
Vasopressin

ASJC Scopus subject areas

Biochemistry
Physiology
Endocrinology
Cellular and Molecular Neuroscience

Access to Document

10.1016/0196-9781(90)90181-4

Cite this

@article{c8c8fe111f1346c5a1cdc5ddb7020e50,

title = "Synthesis and biological activity of a biotinylated vasopressin analog",

abstract = "To study vasopressin receptor-mediated endocytosis using electronmicroscopy methods and to develop avidin affinity columns for receptor purification, we synthesized and tested the biological properties of a biotinylated vasopressin (VP) analog [1-(2-mercapto) propionic acid] 8-[lysine-N6-biotin] VP (B-MLVP). B-MLVP was prepared by coupling biotin to the epsilon amine of the lysine residue in [1-(2-mercapto) propionic acid] 8-(lysine) VP (MLVP). The structure of HPLC purified B-MLVP was confirmed by fast atom bombardment mass spectrometry. B-MLVP effectively competed for arginine vasopressin (AVP) binding sites in canine renal plasma membranes on the surface of LLC-PK1 kidney cells. Dissociation constants of 15 nM and 202 nM were calculated from the results of competition binding assays conducted with membranes and cells, respectively. B-MLVP stimulated adenylate cyclase activity and elevated cellular 3′,5′,cyclic-AMP (cAMP) content in a manner similar to AVP, indicating it is an agonist of VP action in renal tissue. These observations indicate that B-MLVP is an agonist of VP action and may be used to study renal VP receptors by employing avidin coupled to various reporter groups.",

keywords = "Biotin, Biotinylated analog, Vasopressin",

author = "Lutz, {Ward H.} and Londowski, {James M.} and Rajiv Kumar",

note = "Funding Information: The parent peptide, MLVP, was synthesized and purified as previously described (8). B-MLVP was prepared by coupling biotin to the epsilon amine of the lysine residue in MLVP (Fig. 1). This coupling was accomplished by reacting MLVP with 10 molar 1Works upported by NIH Grant DK-25409. 2Dr. W. Lutz was supported by NIH Training Grant DK-07013. 3Requests for reprints should be addressed to Rajiv Kumar, MBBS, Mayo Clinic, 911 Guggenheim Bldg., Rochester, MN 55905.",

year = "1990",

doi = "10.1016/0196-9781(90)90181-4",

language = "English (US)",

volume = "11",

pages = "687--691",

journal = "Peptides",

issn = "0196-9781",

publisher = "Elsevier Inc.",

number = "4",

}

TY - JOUR

T1 - Synthesis and biological activity of a biotinylated vasopressin analog

AU - Lutz, Ward H.

AU - Londowski, James M.

AU - Kumar, Rajiv

N1 - Funding Information: The parent peptide, MLVP, was synthesized and purified as previously described (8). B-MLVP was prepared by coupling biotin to the epsilon amine of the lysine residue in MLVP (Fig. 1). This coupling was accomplished by reacting MLVP with 10 molar 1Works upported by NIH Grant DK-25409. 2Dr. W. Lutz was supported by NIH Training Grant DK-07013. 3Requests for reprints should be addressed to Rajiv Kumar, MBBS, Mayo Clinic, 911 Guggenheim Bldg., Rochester, MN 55905.

PY - 1990

Y1 - 1990

N2 - To study vasopressin receptor-mediated endocytosis using electronmicroscopy methods and to develop avidin affinity columns for receptor purification, we synthesized and tested the biological properties of a biotinylated vasopressin (VP) analog [1-(2-mercapto) propionic acid] 8-[lysine-N6-biotin] VP (B-MLVP). B-MLVP was prepared by coupling biotin to the epsilon amine of the lysine residue in [1-(2-mercapto) propionic acid] 8-(lysine) VP (MLVP). The structure of HPLC purified B-MLVP was confirmed by fast atom bombardment mass spectrometry. B-MLVP effectively competed for arginine vasopressin (AVP) binding sites in canine renal plasma membranes on the surface of LLC-PK1 kidney cells. Dissociation constants of 15 nM and 202 nM were calculated from the results of competition binding assays conducted with membranes and cells, respectively. B-MLVP stimulated adenylate cyclase activity and elevated cellular 3′,5′,cyclic-AMP (cAMP) content in a manner similar to AVP, indicating it is an agonist of VP action in renal tissue. These observations indicate that B-MLVP is an agonist of VP action and may be used to study renal VP receptors by employing avidin coupled to various reporter groups.

AB - To study vasopressin receptor-mediated endocytosis using electronmicroscopy methods and to develop avidin affinity columns for receptor purification, we synthesized and tested the biological properties of a biotinylated vasopressin (VP) analog [1-(2-mercapto) propionic acid] 8-[lysine-N6-biotin] VP (B-MLVP). B-MLVP was prepared by coupling biotin to the epsilon amine of the lysine residue in [1-(2-mercapto) propionic acid] 8-(lysine) VP (MLVP). The structure of HPLC purified B-MLVP was confirmed by fast atom bombardment mass spectrometry. B-MLVP effectively competed for arginine vasopressin (AVP) binding sites in canine renal plasma membranes on the surface of LLC-PK1 kidney cells. Dissociation constants of 15 nM and 202 nM were calculated from the results of competition binding assays conducted with membranes and cells, respectively. B-MLVP stimulated adenylate cyclase activity and elevated cellular 3′,5′,cyclic-AMP (cAMP) content in a manner similar to AVP, indicating it is an agonist of VP action in renal tissue. These observations indicate that B-MLVP is an agonist of VP action and may be used to study renal VP receptors by employing avidin coupled to various reporter groups.

KW - Biotin

KW - Biotinylated analog

KW - Vasopressin

UR - http://www.scopus.com/inward/record.url?scp=0025046882&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025046882&partnerID=8YFLogxK

U2 - 10.1016/0196-9781(90)90181-4

DO - 10.1016/0196-9781(90)90181-4

M3 - Article

C2 - 2172938

AN - SCOPUS:0025046882

SN - 0196-9781

VL - 11

SP - 687

EP - 691

JO - Peptides

JF - Peptides

IS - 4

ER -

Synthesis and biological activity of a biotinylated vasopressin analog

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this