High-level and effective production of human mannan-binding lectin (MBL) in Chinese hamster ovary (CHO) cells

Ohtani, Katsuki; Suzuki, Yasuhiko; Eda, Souji; Kawai, Takao; Kase, Tetsuo; Keshi, Hiroyuki; Sakai, Yoshinori; Yamamoto, Satoshi; Sakamoto, Takashi; Wakamiya, Nobutaka

doi:10.1016/s0022-1759(98)00190-2

Cited by 38 publications

(27 citation statements)

References 31 publications

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“…This phenomenon is consistent with findings of previous reports [31][32][33]. So far, only a single peak at a high-molecular-mass position was reported for purified serum MBL in humans [34][35][36][37] and other species [37][38][39]. It is simply thought that low-molecular-mass MBL (corresponding to peak 2) was overlooked because it did not bind to the carbohydrate resin, and only the fractions which did bind to the resin (corresponding to peak 1) were able to proceed to the next step in purification or identification.…”

Section: Discussionsupporting

confidence: 93%

Relationship between gene polymorphisms of mannose‐binding lectin (MBL) and two molecular forms of MBL

et al. 2003

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Mannose-binding lectin (MBL) activates complement through MBL-associated serine proteases (MASP). A deficiency in MBL due to mutations at exon 1 of the human MBL gene is reported to cause vulnerability to infection. We examined sera of known MBL genotype by gel filtration and assessed their elution patterns using an ELISA for MBL and identified two MBL forms, a high-molecular-mass form and a lower-molecular-mass form. By the identification of either or both forms in individual sera, three types of patterns emerged: type 1 consisted of a high-molecular form; type 2, of a low-molecular form; and type 3, of both forms. Types 1, 2 and 3 corresponded, respectively, to a wild type (A/A), a homozygous mutation at codon 54 (B/B) and their heterozygote (A/B). One exception was a heterozygous LXPA/LYPB phenotype that exhibited the type-2 pattern. Binding to mannan and MASP-1/3 occurred exclusively with the high-molecular form. An apparent MBL deficiency does not in fact represent a deficiency in MBL molecules but rather the presence of circulating oligomeric mutant MBL with impaired function.

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Section: Discussionsupporting

confidence: 93%

Relationship between gene polymorphisms of mannose‐binding lectin (MBL) and two molecular forms of MBL

et al. 2003

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“…Compared with the unmodified polypeptide, the MBL chain shows a mass increase that accounts for the posttranslational modifications usually observed in collagens, i.e., hydroxylation of the proline and lysine residues in the Y position of the repeating Gly-X-Y sequence and glycosylation of hydroxylysine residues. Previous analyses have indicated the presence of 3.7-4 hydroxyproline and 3.2-4 hydroxylysine residues per MBL chain (31,32). Our own results are consistent with the presence of 4 hydroxyproline and 4 hydroxylysine residues, among which 3-4 carry the characteristic O-linked glucosylgalactosyl disaccharide observed in other proteins containing collagen-like sequences such as rat MBL (33), human C1q (34), or adiponectin (35).…”

Section: Discussionsupporting

confidence: 91%

The Two Major Oligomeric Forms of Human Mannan-Binding Lectin: Chemical Characterization, Carbohydrate-Binding Properties, and Interaction with MBL-Associated Serine Proteases

Teillet¹,

Dublet

Andrieu³

et al. 2005

The Journal of Immunology

125

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Mannan-binding lectin (MBL) is an oligomeric C-type lectin assembled from homotrimeric structural units that binds to neutral carbohydrates on microbial surfaces. It forms individual complexes with MBL-associated serine proteases (MASP)-1, -2, -3 and a truncated form of MASP-2 (MAp19) and triggers the lectin pathway of complement through MASP-2 activation. To characterize the oligomerization state of the two major MBL forms present in human serum, both proteins were analyzed by mass spectrometry. Mass values of 228,098 ± 170 Da (MBL-I) and 304,899 ± 229 Da (MBL-II) were determined for the native proteins, whereas reduction of both species yielded a single chain with an average mass of 25,340 ± 18 Da. This demonstrates that MBL-I and -II contain 9 and 12 disulfide-linked chains, respectively, and therefore are trimers and tetramers of the structural unit. As shown by surface plasmon resonance spectroscopy, trimeric and tetrameric MBL bound to immobilized mannose-BSA and N-acetylglucosamine-BSA with comparable KD values (2.2 and 0.55 nM and 1.2 and 0.96 nM, respectively). However, tetrameric MBL exhibited significantly higher maximal binding capacity and lower dissociation rate constants for both carbohydrates. In contrast, no significant difference was detected for binding of the recombinant MASPs or MAp19 to immobilized trimeric or tetrameric MBL. As shown by gel filtration, both MBL species formed 1:2 complexes with MASP-3 or MAp19. These results provide the first precise analysis of the major human MBL oligomers. The oligomerization state of MBL has a direct effect on its carbohydrate-binding properties, but no influence on the interaction with the MASPs.

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“…[31][32][33] Early data suggest that administration of purified MBL is safe and may be effective in ameliorating infection frequency in MBL-deficient individuals. 31 Intensive antimicrobial treatment for infection after SCT is often toxic or unsuccessful, and existing strategies to prevent infection such as prophylactic antimicrobials and intravenous immunoglobulin (which contains no MBL) are incompletely effective.…”

Section: Discussionmentioning

confidence: 99%

Mannose-binding lectin gene polymorphisms are associated with major infection following allogeneic hemopoietic stem cell transplantation

Mullighan

Heatley

Doherty

et al. 2002

Blood

179

149

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Life-threatening complications such as graft versus host disease and infection remain major barriers to the success of allogeneic hemopoietic stem cell transplantation (SCT). While pretransplantation conditioning and posttransplantation immunosuppression are important risk factors for infection, the reasons that similarly immunosuppressed transplant recipients show marked variation in frequency of infection after allogeneic SCT are unclear. Mannose-binding lectin (MBL) deficiency is a risk factor for infection in other situations where immunity is compromised. We investigated associations between MBL2 gene polymorphisms and risk of major infection following allogeneic SCT. Ninety-seven related allogeneic donor-recipient pairs were studied. Clinical data including survival, days of fever, graft versus host disease incidence and severity, and infection were collected by case note review. Five single-nucleotide polymorphisms in the MBL2 gene were genotyped using the polymerase chain reaction and sequence-specific primers. MBL2 coding mutations were associated with an increased risk of major infection following transplantation. This association was seen for donor (

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High-level and effective production of human mannan-binding lectin (MBL) in Chinese hamster ovary (CHO) cells

Cited by 38 publications

References 31 publications

Relationship between gene polymorphisms of mannose‐binding lectin (MBL) and two molecular forms of MBL

Relationship between gene polymorphisms of mannose‐binding lectin (MBL) and two molecular forms of MBL

The Two Major Oligomeric Forms of Human Mannan-Binding Lectin: Chemical Characterization, Carbohydrate-Binding Properties, and Interaction with MBL-Associated Serine Proteases

Mannose-binding lectin gene polymorphisms are associated with major infection following allogeneic hemopoietic stem cell transplantation

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