Structural studies on a glycoprotein isolated from alveoli of patients with alveolar proteinosis

Bhattacharyya, Sambhu N.; Sahu, Saura C.; Lynn, William S.

doi:10.1016/0005-2795(76)90288-9

Cited by 68 publications

(22 citation statements)

References 28 publications

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“…The protein was found to be rich in glycine and had characteristics similar to that previously described for rat and dog glycoproteins A (1 1). There are also similarities between the glycoprotein composition presently reported and that of alveolar glycoprotein Mr= 36,000 previously described by Bhattacharyyra and Lynn (14) who previously demonstrated a high glycine content and collagenase-like sequences in the alveolar proteinosis glycoprotein. Because of these similarities in size and composition, comparisons of the protein from human alveolar proteinosis lavage and the glycoprotein A from normal human lavage were made.…”

Section: Fig 5 Purified Glycoproteins a After Separation By Isoelecsupporting

confidence: 84%

“…Similar glycoprotein has been identified in lung lavage fluid obtained from patients with alveolar proteinosis (1 4). Bhattacharyya and Lynn (14) previously characterized surfactant-associated glycoproteins and provided data that the molecules migrating with Mr = 30,000-40,000 are fragments of a larger Mr = 80,000 dalton polypeptide and have recently described complex N-linked carbohydrates on these molecules (1 5). Several distinct proteins reactive with antisera generated against glycoprotein A have been identified with characteristics of glycoproteins A in lung lavage fluid (16).…”

Section: -D-ief-page Two-dimensional Ief Page Ief-sds-page Isoelectmentioning

confidence: 99%

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Characteristics of Human Surfactant-Associated Glyoproteins A

et al. 1985

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ABSTRAm. Surfactant-associated glycoprotein A [molecular weight (Mr) = 34,000, isoelectric point (PI) 4.6-5.01and its sulfhydryl dependent oligomers were purified and partially characterized from surfactant obtained from human alveolar lavage. Two major forms of the protein were identified by silver stain and immunoblot analysis of surfactant using human surfactant-associated glycoprotein A antisera: glycoprotein A2, M r = 34,000 and glycoprotein A], Mr = 28,000. The larger form was reduced to Mr = 28,000 by treatment with endogylcosidase F, indicating the presence of complex N-linked oligosaccharide on the molecule. Charge heterogeneity was decreased and the isoelectric point increased-by treatment with neuroaminidase, supporting the presence of sialic acid. Homology between the proteins Mr = 34,000 and 28,000 was confirmed by analysis of two-dimensional tryptic and chymotryptic peptides of 1251-iodo-glycoproteins Al and A2 which were identical. The protein was very rich in glycine and its amino acid composition was similar to that of glycoprotein A previously reported for the dog and rat. Treatment of glycoproteins A with bacterial collagenase resulted in the generation of highly glycosylated peptides PtZr = 20,000-22,000, pI 4.6-5.0, which no longer formed sulfhydryldependent oligomers, supporting the presence of significant collagen-like region in the molecule. In the absence of reducing agents, glycoprotein A from surfactant was present as sulfhydryl-dependent dimers and larger oligomers. Higher molecular weight aggregates of glycoproteins A were also present in lavage material even after sulfhydryl reduction. Glycoproteins A were identified in surfactant from amniotic fluid, normal adult lung lavage, human cadaver lung lavage, and material obtained from lung lavage from a patient with alveolar proteinosis. Alveolar proteinosis proteins contained larger amounts of the higher molecular weight aggregates and smaller molecular weight proteolytic fragments of glycoproteins A than material obtained from other sources. Peptide mapping of the '251-iodinated aggregates, approximately Mr = 50,000,70,000, 100,000, and greater were identical to glycoproteins A (Mr = 34,000 and 28,000) from normal human lung lavage. A smaller immunoreactive form (Mr = 20,000) shared several peptides but lacked others, supporting its origin as a proteolytic fragment of glycoproteins A1 or Az. Human glycoprotein A2 is a complex N-linked glycoprotein likely representing the glycosylated form of a polypeptide precursor Mr =

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Section: Fig 5 Purified Glycoproteins a After Separation By Isoelecsupporting

confidence: 84%

Section: -D-ief-page Two-dimensional Ief Page Ief-sds-page Isoelectmentioning

confidence: 99%

Characteristics of Human Surfactant-Associated Glyoproteins A

et al. 1985

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“…1 (27), in a volume of 0.5 ml buffer was applied to the top ofthe column, and fractions of0. 5 (30), using 3 X 7-cm minislab gels. 1% fl-mercaptoethanol was added to samples where indicated as a disulfide reducing agent.…”

Section: Methodsmentioning

confidence: 99%

“…The apoproteins present in this complex have been the subject of many studies over the past few years which have helped to elucidate the physiologic role they play in surfactant activity. While initial interest seemed focused primarily on a 35,000-D major glycoprotein (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17), more recent studies have described hydrophobic 5,000-18,000-D apoproteins (18)(19)(20)(21)(22)(23)(24)(25)(26)(27) Revak. Receivedfor publication 25 June 1987 and in revisedform 28 September 1987. expression of surfactant surface tension reduction activity.…”

Section: Introductionmentioning

confidence: 99%

Use of human surfactant low molecular weight apoproteins in the reconstitution of surfactant biologic activity.

Revak

Merritt²,

Degryse³

et al. 1988

J. Clin. Invest.

134

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Two low molecular weight (LMW) apoproteins were isolated from human pulmonary surfactant. SDS polyacrylamide gel analysis showed one protein (SP 18) to have an apparent molecular weight of 18,000 when unreduced and 9,000 D after reduction. The second protein (SP 9) migrated at -9,000 D in the presence or absence of reducing agents. Both proteins contain a high number of hydrophobic amino acids. The NH2-terminal sequence of SP 18 was determined to be: NH2-phe-proile-pro-leu-pro-tyr-. A cDNA clone isolated from a human adult lung cDNA library contained a long open reading frame encoding at an internal position the human SP 18 amino-terminal sequence.Mixtures of phospholipids (PL) and SP 9 and SP 18 were assessed for their capacity to reduce surface tensions on a pulsating bubble surfactometer. The addition of 1% apoprotein resulted in a reduction of surface tension after 15 s from 42.9 dyn/cm for PL alone to 16.7 and 6.3 dyn/cm for preparations containing SP 9 and SP 18, respectively. In vivo assessment of reconstituted surfactant activity was performed in fetal rabbits. Reconstituted surfactant consisting of PL + 0.5% SP 18 instilled intratracheally at delivery resulted in a marked increase in lung compliance, while the incorporation of 0.5% SP 9 yielded a moderate increase. These data show the ability to produce biologically active surfactant by the addition of isolated LMW apoproteins to defined PL.

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“…Bhattacharyya et al (38) have described short regions of Gly-Xaa-Yaa repeats in this protein. However, there is no homology between the sequence of the canine PSAP and the published sequences of small segments of the alveolar proteinosis glycoprotein (37).…”

mentioning

confidence: 99%

Structure of canine pulmonary surfactant apoprotein: cDNA and complete amino acid sequence.

Benson¹,

Hawgood²,

Schilling³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

151

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The apoproteins of pulmonary surfactant (PSAP) are thought to be critical for normal surfactant function. They bind to surfactant phospholipids and enhance their ability to form surface ifims in vitro. These acidic glycoproteins have monomeric molecular weights of 36,000, 32,000, and 28,000 . Each member of this family of proteins has a similar amino acid composition and their differences in electrophoretic mobility are due in part to glycosylation. We have derived the full amino acid sequence of PSAP-32 from the nucleotide sequence of PSAP cDNA. A cDNA library was prepared from canine lung poly(A)+ RNA and screened with oligonucleotide probes that were based on the NH2-terminal amino acids of PSAP-32 determined by Edman degradation. This protein has the striking feature of collagen-like and non-collagen-like sequences in the same polypeptide chain. There are 24 Gly-Xaa-Yaa triplets, where Yaa is often hydroxyproline. These repeats comprise one-third of PSAP near the NH2 terminus. The remaining two-thirds of PSAP is resistant to bacterial collagenase digestion and contains a possible N-glycosylation site near the carboxyl terminus. The NH2-terminal one-third of PSAP-32 probably contains the cysteine involved in interchain disulflde bonds.Lung surfactant is a mixture of lipids and proteins in part responsible for the maintenance of alveolar stability by lowering the surface tension at the air-liquid interface (1). To decrease the interfacial tension, lung surfactant must be able to rapidly form a surface film. Previously, King and Clements (2) demonstrated that protein-depleted canine surfactant lipids formed surface films more slowly than the intact surfactant containing about 10% protein. They showed that two proteins (Mr 34,000 and 10,000) in canine lung surfactant were lung-specific (3). While the smaller of these proteins has not been extensively investigated, acidic glycoproteins of Mr 28,000-40,000 have been identified in the lung surfactant from several species (4-11).The ability of isolated canine surfactant proteins of Mr 28,000-40,000 to enhance the rate of surface film formation in vitro has been investigated. These proteins readily associate with synthetic phospholipids (12) and extracted surfactant lipids (13,14) and in the presence of calcium ions promote extremely rapid formation of phospholipid surface films (14). Calcium ions are also required for the rapid spreading of native canine surfactant (13). These results have suggested a critical role for pulmonary surfactaInt apoprotein (PSAP) in normal lung function.Canine PSAP comprises a family of proteins of Mr 28,000, 32,000, and 36,000 (PSAP-28, -32, and -36) (15). They have the same NH2-terminal amino acid, isoleucine, and have nearly identical amino acid compositions. All three contain large quantities of glycine (17 mol %) and proline (10 mol %). Approximately half of the prolines are hydroxylated. These proteins are variably glycosylated. Removal of the carbohydrate from PSAP-32 and PSAP-36 by endoglycosidase F digestion results in p...

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Structural studies on a glycoprotein isolated from alveoli of patients with alveolar proteinosis

Cited by 68 publications

References 28 publications

Characteristics of Human Surfactant-Associated Glyoproteins A

Characteristics of Human Surfactant-Associated Glyoproteins A

Use of human surfactant low molecular weight apoproteins in the reconstitution of surfactant biologic activity.

Structure of canine pulmonary surfactant apoprotein: cDNA and complete amino acid sequence.

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